Gene
EditingServices
Comprehensive CRISPRSolutions from Screening
to Clinical Trials「2023-2024 Edition」

www.genscript.com

Make People and Nature HealthierThrough Biotechnology

ABOUT US
GenScript Biotech Corporation (stock code: HK.1548) is a leading global provider of life
science research, development and manufacturing services. Rooted in solid gene synthesis
technology, GenScript has established four major platforms: life science service and productplatform, biomedical contract development manufacture organization (CDMO) platform, celltherapy platform and industrial synthetic biological products platform.
Founded in 2002, GenScript established its R&D and manufacturing headquarters in Nanjing,
China in 2004. In 2015, GenScript was listed on the Main Board of the Stock Exchange ofHong Kong, with legal entities in the United States, China, Hong Kong, Japan, Singapore, theNetherlands and Ireland. It operated business in over 100 countries and regions worldwide,
providing quality, convenient and reliable services and products for more than 100,000
customers.
As of Jun 30th 2022, GenScript owned more than 5,500 employees worldwide, with over 38%
of them holding a Ph.D. or master's degree. GenScript has a number of intellectual property
rights, including more than 190 granted patents and more than 820 patent applications, aswell as a high dense technical secrets. With its mission of “making people and nature
healthier with biotechnology” , GenScript is committed to be one of the most trusted
biotechnology companies in the world. As of Jun 30 th 2022, GenScript’ s services andproducts have been cited in over 74,700 peer-reviewed international academic periodical
articles.

HISTORY & MILESTONES OFGENSCRIPT GROUP

2002

2014

2017

Founded in

Founded Legend Biotech

Legend Biotech and Janssen

New Jersey, US(Cell Therapy Segment)

entered into global strategic
collaboration on cilta-cel

2013

2015

Established Bestzyme

GenScript was listed on HKEX

(Industrial Synthetic Biology(stock code: HK.1548)

Product Segment)

2020

2022

Legend Biotech was listed on Nasdaq

CARVYKTI® granted approval by(NASDAQ: LEGN)US FDA, EC and Japan MHLW

Launched GenScript ProBio

2018

2021

Established Biologics

Group, ProBio and Legend Biotech

CDMO Segment

received funding of(GenScript ProBio)$1 billion from Hillhouse CapitalContent

01
02
03
04
05

Introduction to Genome Editing Service
Technology Platform

01

Production Capacity02

Service Advantages

03

Genome Editing Reagent Service
Synthetic sgRNA

05

CRISPR HDR Knock-in Templates

07

CRISPR Non-viral CAR-T Knock-In Optimization Kits

16

cGMP sgRNA & HDR Knock-in template

17

CRISPR gRNA Library

19

GenCRISPR™ gRNA/Cas9 Plasmid Construction

24gRNA/Cas9 Lentivirus Packaging Service

27

Genome Editing Model Construction Service
GenCRISPRTM Gene Editing Cell Lines

31

GenCRISPR™ Microbial Genome Modification

39

Resource Center
Bioinformatics Tools

41

Technical Guidance Materials

42

Articles Published by Customers

43

Customer Feedback

46

Order Method and Order Query
Order Method

48

Order Query48

01
Introduction to
Genome Editing Service

Technology Platform.....................................................................01
Production Capacity..................................................................... 02
Service Advantages.......................................................................03

Introduction to Genome Editing Service

Technology Platform
GenScript provides complete gene editing solutions, supporting target gene screening and sorting,plasmid/lentivirus/ RNP multiple delivery system experiments, and KO/KI cell line or animal model engineering.

Step1 Target gene screening

Step3 Drug development

GenCRISPRTM
Genome Editing Cell Lines

CRISPR gRNA Library

Step2 Genome editing model constructiongRNA

HDR template

CRISPR gRNA plasmidgRNA/Cas9 Lentivirus package Chemical synthesis sgRNAHot

HDR knock-in templatesHot

Phase-appropriate material and comprehensive QA/QC documentation for successful IND filing

Discovery
Application

Screening for GOI
In vitro functional assays
Small scale in vivo validation

Preclinical Development
Animal Studies/
Efficacy

Clinical Trials
IND
FilingSafety/Toxicology

ISO 9001
QMS

GMP-Like
cGMP

Solutions01

RUO to cGMP sgRNA/ssDNA/dsDNA
Enabling Seamless Transition From Research To Clinics

Phase I

Phase II/III

Introduction to Genome Editing Service

Production Capacity

ISO 9001 Certified

P2 Laboratory: complying with

Provincial demonstration intelligent

the new Biosecurity Act

workshop: high-throughput & efficiency

CRISPR nucleic acids production platform

Synthesis

Quality Control

Automatic Labeling

Automatic Weighting
and Distributing

Purification

Lentivirus Packaging and Cell line Development Platform

BSL2 Level Biosafety Cabinet

Flow Cytometer

Fluorescence Microscope

02

Introduction to Genome Editing Service

Service Advantages

Diverse Technology Platforms

Excellent Production Capacity

High-throughput & large scale platform form
μg to g level deliverable
Support screening, validation to GMP request

Chemical synthesis of sgRNA up to 180 nt
Expert in different sequence & highproportion modification

Strict Quality Control Standards

Professional Technical Support

Customize QC indicators based on
application Improve application successrate and stability

Project management team led by Ph.Ds.
Timely solution of production andapplication problems

Please scan the QR code to visit CRISPR Service home page for more information

GenCRISPRTM Product and Service License
GenCRISPR™ products and services are licensed by Broad Institute, Harvard University, MIT and ERS Genomics in the United States. GenCRISPR™
products and services are protected by US 10,946,108, US 10,930,367, US 10,781,444, US 10,711,285, US 10,577,630, US 9,840,713, US 9,822,372, US
8,999,641, US 8,993,233,US 8,945,839, US 8,932,814, US 8,906,616, US 8,895,308, US 8,889,418, US 8,889,356, US 8,871,445, US 8,865,406, US
8,795,965, US 8,771,945,US 8,697,359 and equivalent patents in many countries, licensed by Broad Institute, Inc. Cambridge, Massachusetts, protectedby US 10,000,772, US 10,113,167, US 10,227,611,US 10,266,850, US 10,301,651, US 10,308,961, US 10,337,029, US 10,351,878, US 10,358,658, US
10,358,659, US 10,385,360, US 10,400,253, US10,407,697, US 10,415,061, US 10,421,980, US 10,428,352, US 10,443,076, US 10,487,341, US
10,513,712, US 10,519,467, US 10,526,619, US10,533,190, US 10,550,407, US 10,563,227, US 10,570,419, US 10,577,631, US 10,597,680, US
10,612,045, US 10,626,419, US 10,640,791, US10,669,560, US 10,676,759, US 10,752,920, US 10,774,344, US 10,793,878, US 10,900,054 andequivalent patents in many countries, and are licensed by ERS Genomics Limited.

03

02

Genome Editing
Reagent Service

Synthetic sgRNA............................................................................. 05
CRISPR HDR Knock-in Templates.............................................07
CRISPR Non-viral CAR-T Knock-In Optimization Kits.........16
cGMP sgRNA & HDR Knock-in template.................................17
CRISPR gRNA Library....................................................................19
GenCRISPR™ gRNA/Cas9 Plasmid Construction................ 24
gRNA/Cas9 Lentivirus Packaging Service.............................27

Genome Editing Reagent Service

Synthetic sgRNA
Traditional CRISPR editing techniques involve transfecting expression plasmids encoding guide RNA and/or Cas9
nuclease into host cells. However, combining Cas9 protein with synthesized sgRNA into ribonucleoprotein (RNP)
prior to delivery can significantly simplify the experimental process, while improving editing efficiency, reducing
off-target effects, and avoiding immune response.
GenScript offers sgRNA synthesis service with guaranteed sequence correctness, which is more efficient, less toxic
and safer than plasmid based editing solutions, improving efficiency and saving time and labor costs for your project.

Service Features

A

B

Chemically synthesized sgRNA

A

VS

B

In vitro transcribed sgRNA

Ready to use and cost-effective

5-6 steps of synthesis work with high cost

Low cytotoxicity, good stability, high editing efficiency

High cytotoxicity, poor stability, low editing efficiency

Accurate sequence with guaranteed deliver quantity

Non-guaranteed success rate

High batch to batch consistency

Large differences among batches

Service Details
Service Name

Length (nt)*Specification (nmol)*

Delivery Time(Business Day)

Delivery standard

EasyEdit sgRNA97-103

1.5-500 nmol

Form 6 days

Accurate sequence
Guaranteed delivery

SafeEdit sgRNA97-1033-1,000 nmol

From 9 days

Accurate sequence
HPLC purification

GenScript provide GMP-Like & cGMP sgRNA to support preclinical development and clinical trials, please find the information in page 17-18.

#

05

Genome Editing Reagent Service

Cases
Case 1: High Editing Efficiency
Editing efficiency of IVT sgRNA and EasyEdit
sgRNA transfected with liposome (HEK293 cells)

Editing efficiency of IVT sgRNA and EasyEdit
sgRNA by electroporation transfection method (HEK293 cells)
100%

Editing Efficiency

Editing Efficiency

70%
60%
50%
40%
30%
20%
10%

IVT sgRNA

0%

EasyEdit sgRNA

Target sequence 1

Target sequence 2

90%
80%
70%
60%

IVT sgRNA
EasyEdit sgRNA

50%

Target sequence 3

Target sequence 1

Target sequence 2

Target sequence 3

After liposome transfection and electroporation transfection, the chemosynthetic EasyEdit sgRNA showed high editing efficiency,
which was consistent with the literature reports[1].

400,000
300,000
200,000
100,000
0
Day1

Day2

Blank control IVT sgRNA

Day3

sgRNA of target sequence 2
500,000
400,000
300,000
200,000
100,000
0

Day4

Day1

EasyEdit sgRNA

Day2

Blank control IVT sgRNA

Day3

Number of living cells

sgRNA of target sequence 1
500,000

Number of living cells

Number of living cells

Case 2: Low Cytotoxicity
sgRNA of target sequence 3
500,000
400,000
300,000
200,000
100,000

Day4

EasyEdit sgRNA

0
Day1

Day2

Blank control IVT sgRNA

Day3

Day4

EasyEdit sgRNA

EasyEdit sgRNA cells showed lower cytotoxicity and less impact on host cell proliferation and cell activity, which was consistent
with the literature reports[2]. IVT sgRNA has 5’ triphosphate modification, which can cause cellular immune response, resulting incytotoxicity and interfere with subsequent experiments, while chemosynthetic EasyEdit sgRNA has no such interference factor.

Case 3: High Stability(Stored at 37 °C for 8 h)

IVT sgRNA

IVT sgRNA

IVT sgRNA(Purified)

IVT sgRNA(Purified)

EasyEdit sgRNA

EasyEdit sgRNA

Positive control

Positive control(Stored at 37 °C for 7 Days)

Chemosynthetic EasyEdit sgRNA is more stable than IVT sgRNA, which degrades during storage (red arrow) because IVT sgRNA
cannot be modified, whereas the chemically synthesized EasyEdit sgRNA can be modified to ensure that sgRNA is morestable in storage and experiments. IVT sgRNA involves a multi-step biological reaction, and there are impurities (blue arrow) suchas buffer and enzyme in the product, while EasyEdit sgRNA is stable in the chemical synthesis process, and has better lot-to-lot
stability than IVT sgRNA, ensuring repeatability and consistency of results in subsequent experiments.
[1] Hendel, A. et al. Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nat.Biotechnol.33 (2015).
[2] Kim et al. CRISPR RNAs trigger innate immune responses in human cells. Genome Res. 28 (2018).

06

Genome Editing Reagent Service

CRISPR HDR Knock-in Templates
One powerful application of CRISPR/Cas genome editing technology involves the precise insertion of DNA
sequences via the homology-directed repair (HDR) pathway. While many factors may affect the efficacy of thisapproach, choosing the right HDR template is certainly an important one. GenScript now offers high quality,
sequence verified HDR templates in both linear and circle format for maximizing the editing efficiency of your CRISPR
experiments, and efficiently support non-viral cell therapy development .

GenExact™ ssDNA
Precise KI and low cytotoxicity

Application

GenWand™ dsDNALong gene knock-in in large scale

Application

Ideal for T cell therapy

Ideal for long gene CRISPR KI

Ideal for cell/animal model
generation

Ideal for screening and scale up
Advantage

Advantage
Minimal cytotoxicity
Precise knock-in, minimized
off-target effectHigh purity and sequence
verified

Specifications
Insertion length: 150-5,000 nt
μg to g scale

Covalently closed ends
protection for better accuracyMore suitable for scale up
High purity and sequence verified
Specifications
Insertion length: 1-10 kbµg to g scale
Research to cGMP grade

GenCircle™ dsDNA
Circular template withno resistance gene

Application
Ideal replacement for plasmid
HDR template
Ideal for scale up and fastdelievery

Advantage
KI efficiency increase by up to30%
429bp backbone, lower
cytotoxicity & higher
transfection efficiency
No antibiotic resistance geneto avoid regulatory concern
High purity and sequence
verified

Research to cGMP grade
Specifications
Insertion length: 1-20 kbµg to g scale
Research to cGMP grade07

Genome Editing Reagent Service

GenExactTM ssDNA Synthesis Service
Linear GenExactTM ssDNA offers the lowest toxicity solution for HDR mediated gene knock-in. Precise insertion ofsequences up to 5kb makes GenExactTM ssDNA an ideal solution for T cell therapy and cell or animal model
engineering.

Why GenExactTM ssDNA?
Cas9/sgRNA complex is cut at the target site,
resulting in double-strand breaks

Genes that obtained precise
knock-in of new DNA fragments
New DNA

ssDNA as a repair template containing
DNA fragments to be knocked in
Low cytotoxicity

Low off-target effect

DNA fragments is knocked-in byhomologous recombination
High knock-in

Improved gene editing

efficiency

accuracy

Service Advantages
Low cytotoxicity and low
off-target effectAccurate sequence guaranteedand high purity

Improve the efficiency and accuracy oftarget gene knock-in

Correct plasmid and Sanger sequencing ofdelivered product

Guarantee the production of cell therapy
products

Deep removal of impurities, with the purity
up to 98%

Powerful synthesis capacity

Support declarations from R&D to clinic

Rich experience in synthesis of sequence
synthesis

Support μg to mg scale delivery

Optimized production technology that is
conducive to scaling up the production

Provide scientific research to cGMP levelproducts

08

Genome Editing Reagent Service

Service Details
Service Type

Length (nt)

Quantity

Production time

Delivery form*

Price

GenExact™ ssDNA#

151-5,000

3μg to 100mg

From 22 days

Lyophilized powder (default)
Liquid solution (customized)

For details,
please consult us

If you need an antisense chain, the cycle remains unchanged. It will be more favorable to order the same sequence again.
*For other length and delivery information's or special difficulty orders needing assessment cycle, please email oligo@genscript.com or call
1-877-436-7274.
#
GenScript provide GMP-Like & cGMP GenExactTM ssDNA to support preclinical development and clinical trials, please find the information in page 17-18.

Cases
The experimental results of Marson Lab at University of California, San Francisco (UCSF) showed that:
GenExact™ ssDNA provides lower cytotoxicity and higher knock-in efficiency than lab-synthesized knock-in
templates
GenExact™ ssDNA of the cGMP scale has a knock-in efficiency of up to 46.2% without the addition of enhancers
Cells that have been knocked-in by GenExact™ ssDNA can reach the patient's dosage (1.5×109), and the
prepared BCMA-CAR cells exhibit tumor-killing activity

GenScript ssCTS

GenScript ssCTS

Internal ssCTS

Internal ssCTS

ssDNA

ssDNA

ssCTS: ssDNA with CTS ( Cas9 target sequences)
Brian R Shy. et.al., bioRxiv(2021)

Brian R. Shy. Et.al. Nature Biotechnology (2022)

09The linear GenWandTM dsDNA double-ended covalently closed design offers improved protection and stability.
GenScript's universal production vector and thermostable enzyme process allows for large-scale manufacturing of long
insert sequences, without the use of animal-derived ingrediants.

Why covalently closed linear dsDNA?
PCR dsDNA

Covalently closed linear
GenWandTM dsDNA

VS

Compared with PCR dsDNA GenWandTM dsDNA has the following advantages:
Professional purification for removing
impurities and low cytotoxicity

Higher stability to avoid digestion anddegradation of exonuclease

Reduced non-homologous end
connection and high knock-in efficiency

Long gene knock-in and cost-effective
scale-up production

Avoid introducing
interference plasmid sequenceService Advantages

Long Gene
Knock-in Template

Scale-up Process
and Production

Strict Quality
Control

Customized
Quality Control

With the synthetic
length of 1-10 kb

Delivery fromμg-scale to g-scale

Strictly control of
impurities and endotoxin

Support R&D, preclinical /clinical researchService Details
Service Type

Length (nt)

Quantity

Turnaround Time

Delivery form*

Price

GenWand™ dsDNA#

1-10 kb

50 μg to g

From 3 weeks

Lyophilized powder (default)
Liquid solution (customized)

For details,
please consult us*For other length and delivery information's or special difficulty orders needing assessment cycle, please email oligo@genscript.com or call
1-877-436-7274.
#
GenScript provide GMP-Like & cGMP GenWandTM dsDNA to support preclinical development and clinical trials, please find the information inpage 17-18.

10

Genome Editing Reagent Service

GenWandTM dsDNA Synthesis Service

Genome Editing Reagent Service

Cases
High purity to avoid unexpected interference
As shown in the case below, a single electrophoretic band and a purity of > 99% can be seen in the GenWand™
dsDNA product inspection.

Purity > 99%
Sample: 8.4 kb dsDNA

10.0 kb
8.0 kb
6.0 kb
5.0 kb
4.0 kb
3.0 kb
2.0 kb
1.0 kb
0.5 kbLane 1: 8.4 kb dsDNA
Lane M: 10000dsDNA marker

High gene knock-in efficiency
The knock-in efficiency of the 2.5 kb sequence* knocked into the RAB11A site of HEK293T cells was measured by the
Neon® Transfection System with 2 μg of dsDNA template, and the gene knock-in efficiency of GenWandTM dsDNA
was higher than that of PCR dsDNA prepared in laboratory (34.90% vs 23.48%).

* The length of knock-in sequence was 2.5 kb (excluding homologous arms)

11

Genome Editing Reagent Service

GenCircleTM dsDNA Synthesis Service
Micro vector with no antibiotic resistance gene & Only 429 bp backbone.
A safe & efficient Knock-in template, transposon vector, viral packaging plasmid or non-viral GOI vector.

Advantages
GenCircleTM dsDNA Win!

VSStandard Plasmid
CpG island

R6K
Ori

KanR

Gene of Interest

AmpR
Gene of Interest

Safer! No Antibiotic Resistance GeneNo antibiotic resistance gene & antibiotic residual
Prevent plasmid replication outside of the engineered
production host strain

Smaller! High fidelity 429 bp backbone
Higher gene expression from equivalent plasmid
quantity
Achieve higher transfection efficiency

Simpler! Lower Cytotoxicity & Immunogenicity
Eliminate CpG islands and other bacterial origin
sequences
Increase stability for longer persistenceMore Efficient! Increase KI/viral packaging/
GOI expression efficiency
Increase knock-in efficiency by up to 30%
Improve viral titer up to 3 folds
Increase transcription levels up to 4 folds, and protein
expression levels up to 39%

Applications Field

Cell Therapy

Gene Therapy

VaccineDisease Model
Gene Function Study

CRISPR KI templateTransposons vector

AAV/LV packaging plasmid
non-viral GOI vector
mRNA preparation template

Antigen expression vector for
DNA vaccine
mRNA preparation template

CRISPR KI templateTransposons vector

12Genome Editing Reagent Service

Service Details
Service

Grade

Insertion
Length*Quantity*

FormatTAT#

Application
Application
Transposons / Transposase
vectorHighPure
GenCircleTM
dsDNA

100 μg - g

1-20 kb

Freeze-dried
powder/Solution liquid(Buffer is
optional)

From 11calendar days

Viral packaging plasmid
GOI transcription & expression

CRISPR KI template

UltraPure* For other insertion length & quantity request, please email oligo@genscript.com or call 1-877-436-7274.
#
This TAT does not include gene synthesis turnaround time.

QC Item

HighPure

UltraPure

Sanger Sequencing

Correct sequence√√

Restriction Analysis

Conforming to reference pattern√√

Spectrophotometer

Delivery quantity guarantee√√

Nanodrop UV Value

A260/280 = 1.8-2.0√√

Electrophoresis

Non-detectable RNA/genome DNA residual√√

Homogeneity≥ 90% Supercoiled√√< 0.01 EU/μg√

TAL Endotoxin
Quantify

13

QC Standard√≤ 0.005 EU/μg

Bioburden TestingNo growth on agar plate after 48 hours

pH

8.0±0.5 (in TE buffer)√

Residual E. coli DNA

Quantitative PCR ≤ 5%√

Residual Host Protein

HCP ELISA ≤ 1%

√

√

√

Case 1. GenCircleTM dsDNA as Knock-in template
Result: CRISPR KI efficiency increased by up to 30%, transposon integration efficiency increased by up to 15%, compared
to standard plasmid.
CRISPR KI template: 2.5kb insert (CD19 CAR+GFP) into TRAC locus of T cell via electrorotation
GenCircleTM dsDNA as CRISPRKI Template
R6K
HA

HA

Knock-in Efficiency (%FITC+)
40.00%

36.43%

10 days

30.00%

GenCircleTM
dsDNA

pUC
20.00%

18.19%

pTemplate
10.00%
0.05%
00.00%
G OI

NC

5.24%

5.91%

pUC

pUC GenCircle
2 μg
Enhancer

GenCircle
2 μg
No enhancer

Transposon vector for KI: 4.8kb insert containing GFP into 293T cell via electrorotation
GenCircleTM dsDNA as
Transposon Vector
R6K

tta
a

pTrans

G OI

15 days
22.00%

R
IT

ITR

a
tta

PiggyBac Knock-in Efficiency (%FITC+)
30.00%
20.00%

GenCircleTM
dsDNA

pUC
12.44%

10.00%
00.00%

7.28%
0.38%

2.99%

NC

pUC

GenCircle

pUC GenCircle

0.5 μg Tn & PBase

1 μg Tn & PBase

Case 2. GenCircleTM dsDNA as virus pacgking plasmid
Result: Viral titer improve up to 3 folds and avoid resistance gene compared to standard plasmid, with stable ITR.
GenCircleTM dsDNA as lentivirus packing plasmid

LT
R

R6K

pLentiG OI

Name

Virus Titer(IFU/mL)

Antibiotic
Resistance GenepRRL-PGK-EGFP(Regular Plasmid)

1.23E+8Significant residue(Ct value<30)

pRRL-PGK-EGFP MF(GenCircleTM dsDNA)

3.81E+8

Undetectable(Ct≥35)

Name

Virus Titer(copies/mL)

Antibiotic
Resistance Gene

CMV EGFP AAV2
(Regular Plasmid)

1.12E+122%

CMV EGFP MF(GenCircleTM dsDNA)

1.22E+12

Undetectable

GenCircleTM dsDNA as AAV packing plasmid
R6K

pAAV2

G OI

14

Genome Editing Reagent Service

Application Case Studies

Genome Editing Reagent Service

Gene transcription level in serum (fM)s
GenCircleTM dsDNA as nonviral vector

R6K

Non-viral
Gene Therapy
Vector

GOI

In Vitro

In Vivo

GOI Transcription Level

800
600

BeforeAfter-9h

400
200
100
80
60
40
20
0
Plasmid 1

Plasmid 2

Plasmid 3

Gencircle
dsDNA

Case 4. GenCircleTM dsDNA as GOI vector for expression
Result: protein expression level increased by up to 39%, with reduced immune response compared to standard plasmid.
GFP Expression
120.00%100.00%
1000

1500

2000
chi…

CMV enhancer

2500

Day 2

80.00%

97.75% 98.55%
73.27%
59.39%

60.00%GFP

CMV promoter

Day 1

SV40 poly(A)signal40.00%
20.00%
0.00%
No EP

0.1 pmol
Plasmid

0.1 pmol
GenCircle dsDNA

Successful Delivered Cases
GenScript has successfully delivered multiple orders with a wide range of GOI lengths, including multiple complex or
difficult-to-synthesize projects.
3.8 kb(AAV transfer plasmid with ITRs,
ITR stable)M

1

2

Lane M : KB Ladder Marker
Lane 1 : U33XXXXX90-1 plasmid
Lane 2 : U33XXXXX90-1 plasmid
digested by EcoRV

15

5.1 kb
M

1

14 kb
2

Lane M : KB Ladder Marker
Lane 1 : U72XXXXX90-1 plasmid
Lane 2 : U72XXXXX90-1 plasmid
digested by Ahdl

M

1

2

Lane M : KB Ladder Marker
Lane 1 : plasmid
Lane 2 : plasmid digested by Xbal

GenScript is proud to offer CRISPR CAR-T knock-in optimization kits to efficiently optimize your CAR insertion processwith pre-validated sequences, design and protocols. Save time on optimization by visually confirming the success of
your next CAR-T knock-in experiment.

Knock-In Optimization Kit Includes
Synthetic SafeEdit sgRNA
HPLC purified sgRNA targeting TRAC or Rab11a for
CAR insertion in T cells

HDR TemplatesHigh Purity Cas9 Proteins
Available in both wild type (Cas9) andenhanced specificity versions (eSpCas9)

Optimized protocol

GenExact™ ssDNA or/and GenWand™ closed-end
dsDNA encoding GFP for easy KI detection

Comprehensive protocol from cell culture, KI experiment,
to editing efficiency detection

Available with Cas9 Targeting Sequence (CTS)
design*Trouble shooting FAQs

How to Build Your Kit?
1. Select TRAC or/and Rab11a as insertion site
2. Choose Cas9 or eSpCas9
3. Choose a single or/and double strand DNA template (ssDNA or/and dsDNA) encoding GFP
4. Select whether include the Cas9 Targeting Sequence (CTS) in HDR Templates for improved knock-in efficiency
5. Download and follow the knock-in optimization protocol and visualize your result
6. Get your kit done! 1.5 nmol sgRNA + 200 μg Cas9 nuclease + 50 μg HDR Templates / per kit

How does the Kit benefit your study?

Save 75% cost and delivery time

Accelerate your study by efficiently

compared to customized reagentsfind out the optimized experiment conditions* Shy, et al. Hybrid ssDNA repair templates enable high yield genome engineering in primary cells for disease modeling and cell therapy manufacturing.
bioRxiv 2021.09.02.458799.

16

Genome Editing Reagent Service

CRISPR Non-viral CAR-T Knock-In Optimization Kits

Genome Editing Reagent Service

cGMP sgRNA & HDR Knock-in template
GenScript is proud to offer current Good Manufacturing Practice (cGMP) manufacturing of synthetic single guide
RNA (sgRNA) and HDR template (ssDNA/dsDNA) supporting non-viral gene and cell engineering development.Our
State-of-the-Art Production Facility operates dedicated cGMP production lines to reliably provide CRISPR sgRNAwith the necessary documentation required for successful IND submission and clinical trials.
Partner with GenScript to accelerate your therapeutic pipeline from early-phase research to clinical applications.

State-of-the-Art
Production Facility

Comprehensive QA/QC
Documentation

Trusted Partner from
Bench to Clinic

cGMP manufacturing of 30mg up
to gram quantities per batch

10+ QC options supporting
global IND filing needs

Successfully delivered
100+ cGMP batches

Compliant with
FDA/EMA/PMDA/NMPA regulations

US Type II DMF and/or CMC
filing support

6 IND approvals

cGMP Production Workflow
Information
Exchange & Design

Pilot study &Process Development

QC & quality
research

Aseptic Fill/Finish& Aliquot

Raw Material
Preparation &Specification

Sequence
Synthesis

Purification

Experienced team successful support

100+

cGMP batches product30

IND filling projects

6

IND approval

GenScript is further expanding cGMP sgRNA & HDR templateproduction capacity to reduce the wait time for our partners.
GenScript’s new Zhenjiang, China 400,000 sq.ft. cGMP facility has
been put into production.

17

Genome Editing Reagent Service

Why GenScript?

Comprehensive QA/QC &Documentation

Qualified Production Facility

cGMP production environment

Manufacturing Summary Report, TSE/BSE Statement

Clean suite with class A isolator in a class C background
Sterile filtration and aseptic fill finish

Batch Records based on Master Batch Record and Change
Control System

Process development and optimization

Identity, purity, sterility, etc. clearly defined

CQA/CPP confirmed

cGMP-compliant material and supplier management

Flexible batch scale from 30 mg-1 g

Validated QC testing procedures

Sufficient capacity! No need to wait for cGMP schedulingLong-term stability testing under various solutions andstorage conditions
Established QMS for training, process deviations, and CAPA

One Stop Solution Support Gene and Cell Therapy Development
Phase-appropriate material and comprehensive QA/QC documentation for successful IND filing
Discovery
Application

Screening for GOI
In vitro functional assays
Small scale in vivo validation

Preclinical Development
Animal Studies/
Efficacy

Safety/Toxicology

IND
FilingClinical Trial
Phase I-Ⅲ

NDA Filing

Commercialization

ISO 9001
GMP-Like

QMS

cGMP
SolutionsRUO to cGMP sgRNA/ssDNA/dsDNA
Enabling Seamless Transition From Research To Clinics

Successful Cases
Bioheng Biotech Co., Ltd.

BRL Medicine Inc.

cGMP sgRNA support the first UCAR-T IND
approval in China

cGMP sgRNA support the first nonviral CAR-T IND
approval globally

Marson Lab

T-Maximum Biotech

ssDNA achieve up to 60% KI efficiency for CAR-T
generation at a GMP-compatible scalecGMP sgRNA supported their MT207 CAR-T celltherapy’s FDA-certified orphan drug designation (ODD)

Contact US
Welcome to consult us about cGMP reagents to support your GCT program at any stage. Professional technical support
manger is at your service.
Email: oligo@genscript.com

Tel: 1-877-436-7274

18

Genome Editing Reagent Service

CRISPR gRNA Library
CRISPR gRNA library is an important tool for high-throughput screening of target genes, which applies the highefficiency and specificity of CRISPR gene editing technology to knock out expressed genes in the genome or activate
gene expression. CRISPR gRNA library construction service applies oligo pool synthesis technology to effectively
reduce the cost and save the time of gene target screening.

Library Construction Workflow

gRNA Design

gRNA Oligo Synthesis

Subcloned toTarget Vector

Library Amplification

Lentivirus Packaging

Service Features
High coverage - NGS sequencing shows that full coverage could be achieved
Excellent homogeneity (90%/10%), Better than the industry level
Provided transfection grade plasmids with endotoxin < 0.005 EU/μg

Service Details
Service Type

Content

Number of gRNAs

Genome-wide gene knockout gRNA library (GeCKO gRNA library)

Human and mouse genome-wide gene knockout

3 or 6 gRNAs/genes

CRISPR transcriptional activation gRNA library (SAM gRNA library)Transcriptional activation in human and mouse
genome-wide

3 gRNAs/genes

Pathway or disease-related gene knockout

Depending on the pathway

ArraygRNA plasmids were synthesized and cloned
separately

Tens to hundreds

Pool

Different gRNA plasmids were mixed together

Up to one hundred thousand

Pathway-focused gRNA library
Customize gRNA library

Application Method
Deep sequencing

Analysis ofremaining sgRNA

Positive
ScreenCRISPRlibrary

Amplify
library

Packagelibrary

Apply virallibrary
Deep sequencing

Negative
ScreenCompare change insgRNA counts
Deep sequencing

Application Method of CRISPR gRNA Library

19

GenScript provides CRISPR gRNA Library (GeCKO v2.0) introduced from Zhang Feng Laboratory of Broad ResearchInstitute. It is patented and can knock out any gene and miRNA in the genome of human or mouse, which has been
widely used in primary human or mouse cells, stem cells, cancer cells and various stable cell lines.
6 different gRNA sequences are designed for each gene in the GeCKO v2.0 library, which is divided into library A andlibrary B. Each library contains 3 different gRNA sequences for each gene and 1,000 non-targeted control gRNAs.
Library A also contains gRNAs targeting miRNAs (each miRNA has 4 targeted gRNAs). In addition, GeCKO v2.0 libraryhas upgraded lentiviral vectors for transduction, which can enrich viruses more efficiently and improve the transfection
efficiency of viruses in primary cells.
To ensure complete knockout for each gene, both library A and library B can be purchased for mixed use. If cell samples
and follow-up high-throughput screening capacity are limited, it is recommended that only library A be used for sorting.

Library Details
Library Type
Human Library A

Sequence Information
65,383 sequences:
19,050 genes were targeted, and each gene had 3 targeted gRNAs
1,864 miRNAs were targeted, and each gene had 4 targeted gRNAs
1,000 control (non-targeted) gRNAs
58,028 sequences:

Human Library B

19,050 genes were targeted, and each gene had 3 targeted gRNAs
1,000 control (non-targeted) gRNAs
67,405 sequences:
20,611 genes were targeted, and each gene had 3 targeted gRNAs
1,175 miRNAs were targeted, and each gene had 4 targeted gRNAs
1,000 control (non-targeted) gRNAs

Mouse Library A

62,804 sequences:
Mouse Library B

20,611 genes were targeted, and each gene had 3 targeted gRNAs
1,000 control (non-targeted) gRNAs

VectorSpecification

pLentiCRISPR v2
pLentiGuide-PuropLentiCas9-Blast
pLentiCRISPR v2
pLentiGuide-PuropLentiCas9-Blast
pLentiCRISPR v2

25/100/200 µg

pLentiGuide-PuropLentiCas9-Blast
pLentiCRISPR v2
pLentiGuide-PuropLentiCas9-Blast

Delivery standard
Transfection grade libraries were delivered with 25 μg as a unit, libraries with 100 μg and 200 μg were delivered
with 4 × 25 μg or 8 × 25 μg as a unit, respectively.
We provide statistical summaries of project reports and NGS validation and COA files, as well as complete, usable
NGS raw data if you require.

Application Method
1) Each gRNA in GeCKO v2.0 library was cloned into the optimized lentivirus vector, which could form a high titervirus after co-transfection with Cas9 vector in 293 cells. Finally, the gRNA sequence could be efficiently transcribedin the target cells;2) GeCKO v2.0 libraries with completed virus packaging should be infected with target cells at a low MOI to ensurethat no more than 1 gRNA enters a single cell;3) After transfection, a round of NGS deep sequencing should be performed before sorting to evaluate the expressionof gRNA in cell pools.
4) After the sorting, the second round of NGS deep sequencing was conducted, and the data were analyzed todetermine the gRNA sequences loss or enrichment in the sorting process, so as to deduce the target genes.

20

Genome Editing Reagent Service

GeCKO gRNA LibraryGenome Editing Reagent Service

CRISPR transcriptional activation gRNA library (SAM sgRNA library)
The dCas9 protein lacking cleavage activity was fused with the transcriptional activation element and equipped withthe gRNA sequence targeting the upstream regulatory region of the gene to form the transcriptional activation
complex, which can precisely target the upstream transcriptional activation region of genes and effectively activate
gene expression.
SAM sgRNA library, a gRNA library targeting the whole gene wide, combines with dCas9 and transcriptional
activation protein to up-regulate the whole gene expression level, which can be applied to high-throughput and rapid
gain-of-function screening.
MS2 RNA aptmers

dCas9
sgRNA
VP64

MS2
p65
HSF1

assembled SAM complex

CRISPR/Cas9 SAM complex consists of three parts: gRNA with two MS2 RNA aptamers, MS2-P65-HSF1 activation accessory protein anddCas9-VP64 fusion protein

Library Details
Library Type

Sequence Information

70,290 sequences:
Human SAM Library

VectorSpecification

pLenti sgRNA(MS2)_zeo
pLenti dCas9-VP64_Blast
pLenti MS2-P65-HSF1-Hygro

Targeted 23,430 genes
Each gene had 3 targeted sequences

pLenti sgRNA(MS2)_puro
pLenti dCas9-VP64_Blast
pLenti MS2-P65-HSF1-Hygro

25/100/200 µg

69,225 sequences:
Mouse SAM Library

Targeted 23,439 genes
Each gene had 3 targeted sequences

pLenti sgRNA(MS2)_puro
pLenti dCas9-VP64_Blast
pLenti MS2-P65-HSF1-Hygro

491 control (non-targeted) gRNAs

Delivery standard
Transfection grade libraries were delivered with 25 μg as a unit, libraries with 100 μg and 200 μg were delivered
with 4 × 25 μg or 8 × 25 μg as a unit, respectively.
We provide statistical summaries of project reports and NGS validation and COA files, as well as complete, usable
NGS raw data if you require.

Application Method
1) SAM libraries with virus packaging should be transfected with target cells at a low MOI to ensure that no morethan 1 gRNA enters a single cell;2) After transfection, a round of NGS deep sequencing should be performed before sorting to evaluate theexpression of gRNA in cell pools.
3) After the sorting, the second round of NGS deep sequencing was conducted, and the data were analyzed todetermine the gRNA sequences loss or enrichment in the sorting process, so as to deduce the target genes.

21

Pathway-focused gRNA library is an ideal tool for targeted screening for specific pathways. Applied with gene targets
identified by the Drug Gene Interaction Database, the library can be designed to sort for pathway or disease-related
genes. All gRNA sequences were pre-designed and verified by the Broad Research Institute and can be customized to
be cloned into lentiviral vectors.

Library Details
Pathway

Number of
Genes

Number ofgRNAs

Pathway

ABC Transporter

100

298

Acute myeloid leukemia

45

135

Adult Stem Cells

58

174

Adipogenesis

126

378

Angiogenesis

92

276

Alzheimer's disease

94

281

Apoptosis

68

204

58

174

cAMP & calcium Signaling
Pathway

B cell receptor signaling
pathway91

271

Bladder cancer

37

111

Cell Cycle

84

252

Cardiovascular Disease

91

273

Cytochrome P450

57

171

Chemokine signaling pathway

134

402

Drug Resistance

348

1,044

Chronic myeloid leukemia

57

171

GPCR

280

840

Colorectal cancer

54

162

Hormone Activity

108

324

Diabetes44

132

Ion Channels (Potassium)

67

201

Drug Metabolism

34

102

Number of Number of
Genes
gRNAs

Tumor Metastasis

60

180

Drug transporters

84

252

Nuclear Hormone Receptors

117

351

ES Cell Differentiation

334

1,002

Oncogenes

289

867

Extracellular Matrix & Adhesion

84

252

Tumor Suppressor

720

2,155

Growth Factor

161

483

WNT Pathway

92

276

Histone Modification

259

776

Please scan the QR code to visit CRISPR gRNA Library Service home page for more information

22

Genome Editing Reagent Service

Pathway-focused gRNA Library

GenScript has a high-throughput oligonucleotide synthesis platform based on semiconductor microarray technology
to accurately synthesize single stranded oligonucleotides. After PCR amplification, double stranded gRNA is cloned
into the selected vector to create a sorting library, which is combined with efficient oligo pool to achieve higher virus
titers.
A fully customized gRNA library with complete coverage and homogeneous distribution is provided for CRISPR geneknockout, CRISPRa and CRISPRi.

Cloning
2000

vector

1500

QC

Cleavage

+

1000
500
0

PCR

0

0 1 2 3 4 5 6 7 8 9 10 11 12 13

NGS valldation
synthesized gRNA oligos

Service Features
Coverage of NGS validation > 99%
Various cloning vectors: dual vector or all-in-one CRISPR vector approved by Zhang Feng Laboratory or otherdesignated vectors
Delivery plasmids can be used directly for virus packaging with flexible delivery (from μg to mg scale)No sequence restriction, short delivery cycle within 4 weeks

Case: The coverage and homogeneity of gRNA library were verified by NGS
5000

4000
Sequences

Genome Editing Reagent Service

Customize gRNA library

3000

2000

10% effective read length

44

90% effective read length206

Homogeneity (90%/10%)

4.68

Mean sequencing depth

117

Maximum depth sequencing

1,068

Theoretical gRNA diversity

62,804

Actual gRNA diversity

62,804

1000

0

0

1

2

3

4

5

6

7

8

9

10

11

NGS Reads(log2)

A gRNA library with a theoretical diversity of 62,804 was tested by NGS, and the results showed that the gRNA librarycompletely contained gRNA sequences covering all designs. In addition, the homogeneity (90%/10% ratio) was lessthan 5, indicating that the required gRNA was homogeneously distributed in the library to ensure screening efficiency.
*For gRNA library service, please send an email to gene@genscript.com or call 1-877-436-7274, and our professional technical support team will be atyour service.

23

The CRISPR reagents provided by GenScript are collected from Zhang Feng Laboratory. The CRISPR products andservices provided by GenScript are licensed by Broad Institute in the United States. These include:sgRNA sequences that have been designed and verified by MIT
Multiple vector types including binary vector and all-in-one are available, and resistance screening or methods canbe selected as needed
Empty vector verified by MIT
Various Cas9: eSpCas9, SpCas9, SpCas9 Nickase, SaCas9 and SAM

Service Features

Specific sgRNA Sequence

Genome-wide Database

Professional Design Tool

On-line OrderingsgRNA sequence verified by MIT
Ensure the specificity andefficiency of genome editing

Human and MousegRNA database
Support for SpCas9 andSAM applications

12 species optional
On-target/off-target scoring

Customized sequence
Vector and vector
label optional

Service Details
Customized Cas9 plasmids

Expression System

Sorting label

eSpCas9 plasmids

Plasmid
LentiviralAmp, Puro
Amp, GFP

SpCas9 plasmids

Plasmid
Lentiviral
AAV

Amp
Amp, Puro
Amp, Neo
Amp, GFP

SpCas9 Nickase plasmids

Plasmid
Lentiviral

Amp
Amp, Puro
Amp, GFP

SaCas9 plasmids

AAV

Amp

SAM gRNA plasmids

Plasmid
Lentiviral

Amp
Amp, Zeo

SAM dCas9-VP64 plasmids

LentiviralAmp, Blast
Amp, GFPSAM MS2-P65-HSF1 plasmids

LentiviralAmp, GFP
Amp, Hygro

SAM plasmids(transcriptional
activation plasmids)

To meet your scientific research needs, GenScript provides empty vector service with vector sequences that havebeen verified by MIT. The vectors contain a 17 bp-1.8 kb expressible ligand in place of a customized sgRNA sequence,
which can be modified according to your needs.
Service Items

Expression System

Sorting label

eSpCas9 and SpCas9 Broad Plasmid Collection

Plasmid
LentiviralAmp, Puro
Amp, GFP

SpCas9 Nickase Broad Plasmid Collection

Plasmid

Amp
Amp, Puro
Amp, GFP

SaCas9 Broad Plasmid Collection

AAV

Amp

24

Genome Editing Reagent Service

GenCRISPR™ gRNA/Cas9 Plasmid Construction

Genome Editing Reagent Service

CRISPR/Cas9 Related Vector Specification
Category

VectorExpression Type

Delivery Type

Vector Type

Sorting LabeleSpCas9 Plasmids

eSpCas9-2A- GFP (PX458)

eSpCas9 & gRNA

Plasmid

All-in-one Vector

AmpR
EGFP

eSpCas9 & gRNA

Plasmid

All-in-one Vector

AmpR
PuroR

eSpCas9 Plasmids

eSpCas9-2A- Puro
(PX459) V2.0

eSpCas9 Plasmids

eSpCas9-LentiCRISPR v2

eSpCas9 & gRNA

LentiviralAll-in-one Vector

AmpR
PuroR

SpCas9 Plasmids

pSpCas9 BB-2A-GFP PX458

SpCas9 & gRNA

Plasmid

All-in-one Vector

AmpR
EGFP

SpCas9 & gRNA

Plasmid

All-in-one Vector

AmpR
PuroR

pLentiCRISPR v2

SpCas9 & gRNA

LentiviralAll-in-one Vector

AmpR
PuroR

pLentiGuide-PurogRNA Only

Lentiviral

Dual Vector

AmpR
PuroR

SpCas9 Plasmids

pLentiCas9-Blast

SpCas9 Only

Lentiviral

Dual Vector

AmpR
BsdR
BleoR

SpCas9 Plasmids

pLentiCas9-EGFP

SpCas9 Only

Lentiviral

Dual Vector

AmpR
EGFP

SpCas9 Plasmids

pGS-gRNAgRNA Only

Plasmid

Dual Vector

AmpR

SpCas9 Plasmids

pGS-gRNA-Neo

gRNA Only

Plasmid

Dual Vector

AmpR
NeoR

SpCas9 Plasmids

pSpCas9 PX165

SpCas9 Only

Plasmid

Dual Vector

AmpRgRNA Only

AAV

Dual Vector

AmpR
EGFP
AmpR

SpCas9 Plasmids

SpCas9 Plasmids

SpCas9 Plasmids
SpCas9 Nickase Plasmids

SpCas9 Plasmids

pSpCas9 BB-2A-Puro(PX459) v2.0

pAAV_SpGuide acceptor(PX552)

SpCas9 Plasmids

pAAV-SpCas9 PX551

SpCas9 Only

AAV

Dual Vector

SpCas9 Nickase Plasmids

pSpCas9n BB PX460

SpCas9 Nickase & gRNA

Plasmid

All-in-one Vector

AmpR

SpCas9 Nickase Plasmids

pSpCas9n BB-2A-GFP PX461

SpCas9 Nickase & gRNA

Plasmid

All-in-one Vector

AmpR
EGFP

SpCas9 Nickase & gRNA

Plasmid

All-in-one Vector

AmpR
PuroR

SpCas9 Nickase Plasmids

pSpCas9n BB-2A-Puro(PX462) V2.0

SpCas9 Nickase Plasmids

pLentiCas9n-Blast

SpCas9 Nickase Only

Lentiviral

Dual Vector

AmpR
BsdR
BleoR

SaCas9 Plasmids

pX601_AAV

SaCas9 & gRNA

AAV

All-in-one Vector

AmpR

25

VectorTranscription Activation (SAM)

pSgRNA(MS2)gRNA Only

Plasmid

SAM Multi Vector

AmpR

Transcription Activation (SAM)

pLenti_sgRNA(MS2)_zeo

gRNA Only

Lentiviral

SAM Multi Vector

AmpR
ZeoR
BeloR

Transcription Activation (SAM)

pLenti_dCas9-VP64_Blast

Cas9 Activator

Lentiviral

SAM Multi Vector

AmpR
BlastR
BleoR

Transcription Activation (SAM)

pLenti_dCas9-VP64_GFP

Cas9 Activator

Lentiviral

SAM Multi Vector

AmpR
EGFP
BleoR

Transcription Activation (SAM) pLenti_MS2-P65-HSF1_Hygro

Activator Adapter

Lentiviral

Multi Vector

AmpR
HygroR
BleoR

pLenti_MS2-P65-HSF1_GFP

Activator Adapter

Lentiviral

Multi Vector

AmpR
EGFP
BleoR

Transcription Activation (SAM)

Expression Type Delivery Type Vector Type

Sorting LabelCRISPR gRNA Database
GenScript provides over 20,000 plentiCRISPR v2 plasmids containing gRNA sequences, and all sgRNA sequences
have been verified by MIT. You can find the relevant gRNA by searching for Gene Name, Gene Symbol, or Gene ID
in the database. You can access the database through the following website or by scanning the QR code.

https://www.genscript.com/gRNA-database.html

26

Genome Editing Reagent Service

Category

Genome Editing Reagent Service

gRNA/Cas9 Lentivirus Packaging Service
Lentiviruses, a type of retrovirus, are one of the most popular tools for gene transduction. Lentivirus can introduce
target genes into some difficult-to-transfect cells, such as primary cells, stem cells, or non-dividing cells, thus greatly
improving the efficiency of target gene transduction. Lentiviral vectors also have significant advantages in transienttransfection and stable strain sorting. GenScript offers a lentiviral packaging service with flexible titers andspecifications and delivery time as fast as 2 weeks.

Service Features
Standard and Professional
Technology Platform

Strict Quality Control

Traceable and authorized cell line materials
Third-generation lentivirus vector: serum-freesuspension system

Test standard that higher than the industry level
ELISA/FACS/qPCR/resistance sorting

Cost effective

Fast Delivery Cycle

Save 20% on getting the same amount of virus

Quick turnaround time of 2 weeks, faster than theindustry average delivery time

Service Details
Titer*>107 IFU/ml>108 IFU/ml>109 IFU/ml

Specification

Cycle

QC

2-3 weeks

p24 ELISA detection
Lonza 180 test
FACS/qPCR/resistance sorting (customized QC)

1 ml
2 ml
1 ml
2 ml
0.1 ml0.2 mlCustomers provide sequences for gene synthesis at an additional charge.
In the lentivirus package project delivered by GenScript, the longest package gene fragment reached 6.6 kb andachieved a high titer level.
*For other titers or specifications informations, please send an email to protein@genscript.com.cn or call 400-025-8686 ext. 5821, and our professional
technical support team will be at your service.

27

Genome Editing Reagent Service

Cases
Case 1: Lentivirus infection experiment

Second-generation virus system - GenScript

Third-generation virus system - GenScript

Third-generation virus system - other companies

EGFP lentivirus was packaged with the second-generation virus packaging system of GenScript, the optimized third-generation
virus packaging system of GenScript and packaging system of other companies, respectively. After the determination of titers, they
were adjusted to the same MOI to infect the same amount of 293T cells, and fluorescent photographs were taken 72 hours after
infection. The upgraded GenScript third-generation system had higher target protein expression.

Case 2: GFP virus infection Jurkat MOI testing

MOI

GFP%

Infection time

10

17.1%

72 h

20

26.1%

72 h

40

41.6%

72 h

80

60.1%

72 h

Jurkat cell is a type of T lymphocyte that is difficult to transfect. Jurkat cells were infected with GFP virus packaged with GenScript
third-generation plasmid system and tested to achieve an infection rate of 60.1% at MOI=80.

28

T ran s fe c ti on E ffi c i e n c yCommercial Vector

Cell Line

Name

MOI

GS Optimized Vector

120.00%

CT26

Mouse colony

BAF3

Mouse B cell

EL4

Mouse T lymphoma

RAW 264.7

Monocyte/macrophage-like cells

JurkatHuman acute T cell leukemia

20.00%

K562

myelogenous leukemia cell line

0.00%

NALM6

B cell precursor leukemia cell

THP-1

Acute monocytic leukemia**
100.00%****80.00%**
60.00%

100

RT

62

KA
JU

M
6

K5

4

P1

N
AL

TH

EL

RA

W
26

6

BA

F3

40.00%

4

**

CT
2

Genome Editing Reagent Service

Case 3: High Transfection efficiency of optimized GLV3 vector

MOI: Multiplicity of infection

Case 4: Gene carrying length testing of GenScript lentivirus packaging system
Gene name

Target gene length ( kb )Titer ( x10^8 IFU/ml )Gene 1

3.8

3.43

Gene 2

4.7

3.59

Gene 35.5

1.85

Gene 46.6

2.61

In general, target genes longer than 3,000 bp are difficult to reach high titer levels and even impossible to package. In the lentivirus
package project delivered by GenScript, the longest package gene fragment reached 6.6 kb and achieved a high titer level.
GenScript can guarantee a titer of 107 IFU/ml for conventional genes (moderate GC content, no complex repeats, no cell death andthe length less than 3 kb).

Please scan the QR code to visit gRNA/Cas9 Lentivirus
Packaging Service home page for more information

29

03

Genome Editing Model
Construction Service

GenCRISPRTM Gene Editing Cell Lines..........................31
GenCRISPR™ Microbial Genome Modification.......... 39

20

Genome Editing
Model Construction Service

GenCRISPRTM Genome Editing Cell Lines
CRISPR/Cas9 technology was used to knock-out genes and knock-in the target sequence, resulting in gene editingcell lines that can be stably passaged. In the field of basic research, it can be used for target gene/protein tracing andfunctional research; in the field of cell/gene therapy and drug development, it can be used in immunotherapy target
discovery, drug screening, disease biology/signaling pathway research and others.

Advantages of GenScript
GenScript provides a variety of CRISPR gene editing cell lines. Relying on the advantages of chemically synthesized
long single-stranded gRNA, we can adopt RNP delivery system with high editing efficiency and low toxicity, andsupport multiple delivery systems of lentivirus and plasmid to provide you with gene-editing cell line services
specifying target genes, editing regions and cells.

High Quality Delivery
Cell lines/pools
verified by sequencing
A variety of validation,
optimization, off-target
analysis services

Compliance Qualification

Professional
Technical Service

Rich Experience

P2 Laboratory: complying with
Over 1,000 cell lines of delivery
the new Biosecurity Act
experience worldwide
Broad/ERS authorization: Providing
Excellent handling ability
CRISPR services in complianceof difficult projects

More efficient projectcommunication and promotion
Timely answering questions
by Ph.D. project manager

Genome Editing Cell Line Construction Process

Host cell
preparation
sgRNA synthesis

1

3-4 weeks

Host cell detection
Promoter verification
3 sgRNAs designed foreach gene

31

Cells aretransfected
to form polyclones

2

1-3 weeks

Transfection/virus/RNP delivery:
Multiple options to
optimize transfection
efficiency
Cell pool testing:sequencing toconfirm successful
knock-out or knock-in

Selectingmonoclonal
culture3

1-4 weeks

FACS/Resistancescreening enriched
positive clones
Select monoclonal

Sorting andverification

4

2-3 weeks

100+ monoclonalsequencing
validation to confirm
successful knock-out
or knock-in of thetarget sequence

Cryopreservation
and QC ofcell lines

5

2-6 weeks

Cryopreservation ofcells
Cell viability detection
Mycoplasma
detection
Sequencing toconfirm allelic geneknock-out in cell lines
or target sequenceknock-inService Type

Service Details

Cell Line Options

Deliverables**DeliveryTme

EZ knock-out cellline service

Single-gene
knock-out stable
cell line

90 + common cell lines suitable for transfection(including A549, CHO-K1, HT-29, MDA-MB-231, 4T1, A20,
HCT116, MCF7, MDCK, U937, and RPMI 8866)

2 full-allelic knockout celllines verified by sequencing
1 negative knock-out cellpool control

8-15 weeks

Customized Single-gene or multi-gene
knock-out
knock-out stable cell line
cell line service

Any cancer cell line

1-2 full-allelic knockout celllines verified by sequencing
12-19 weeks
1 negative knock-out cellpool control

Full-length gene Whole gene fragments
knockout
knock-out stable cell line
cell line servicNew

50 + common cell lines suitable for transfection(including A549, CHO-K1, HEK293, HEK293T, HT-29,
MDA-MB-231, 4T1, A20, HCT116, and MCF7)

1 full-allelic knockout celllines verified by sequencing
12-20 weeks
1 negative knock-out cellpool control

Knock-out cellpool service

Single-gene
knock-out cell pool

Any cell line

Customized
knock-incell line service

Insert or modify genes
at any genomic location

Any cancer cell line

Customized
screeningand off-target
effect detectionNew

Genome-wide libraries
or customized libraries
were used to screen
target genes andiGUIDE was used todetect potential
off-target effects

For any cells can be cultured, please consult us.

Cell pool with knock-out
clones verified bysequencing

5-11 weeks

1 knock-in cell lines verified
by sequencing
1 unmodified control cell

14-24 weeks

Analysis report

For details,
pleaseconsult us*Cell lines are generally supplied by the customer, or for an additional fee if provided by GenScript.
**Congruent gene modification cell lines were verified at mRNA or protein levels as required. The project will be updated once every two weeks.

Other customized services
Additional Services

Details

Reverse transcription PCR (RT-PCR)

RT-PCR sequencing was performed to verify that the knock-out clones
inserted deletion markers on CDS at the mRNA level

Western blot

Knock-out clones were verified by WB, and effective antibodies that bind
specifically to target proteins in host cells were required

FACS analysis

Knock-out clones were verified by FACS, and effective antibodies that bind
specifically to target proteins in host cells were required

Promoter activity analysis

The analysis of promoter (Cbh/CMV/EFS) activity was performed in host cells
to optimize the cutting efficiency of gRNA-Cas9

Optimization of transfection method

The analysis of transfection method of difficult-to-transfect cells was
performed to optimize the cutting efficiency of gRNA-Cas9

Off-target analysis

Multiple potential off-target sites were sequenced to identify knock-out clones
and analyze off-target sites at genome level

Additional monoclonal

An another sequenced monoclone was selected from the cell pool and sent tothe customer*For GenCRISPR™ gene editing cell line services, please send an email to protein@genscript.com or call 1-877-436-7274, and our professional
technical support team will be at your service.

32

Genome Editing
Model Construction Service

Service Details

Genome Editing
Model Construction Service

Full-length Gene Knock-out Cell Line Application Field
GenCRISPR™ delivers full-length knock-out cell lines with long fragments of up to 200 kb in multiple copies. Morethan 20 full-length gene knock-out projects have been successfully delivered worldwide, especially for the following
applications to eliminate potential interference factors.

Unknown genome or DNA sequence

Difficult to obtain high specificity antibodies

Non-coding RNAs, novel subtypes/functions of known genes

Nonspecific binding still exists after routine gene knock-out

Special requirements from regulators

Core signal pathway, important data research

Negative control cell lines for diagnostic kits

Full-length gene knockout eliminates possible interference factors

Gene Knock-in Cell Line Application Field
GenCRISPR™ technology provides congruent gene knock-in cell lines verified by sequencing with high editing
efficiency and more efficient and rapid delivery. To date, we have successfully delivered dozens of projects, including
SHSY5Y, hTERT-RPE1, KM12, HeLa, HCT116, Caco2, C2C12, U2OS, Min6, HepG2, and HEK293, providing support for
the following applications.
TC

15,000

Positiveclone
linkerParentalcell

GFP protein

SNP Research

33

Precise Gene Overexpression

Tracer Gene LabelingGenome Editing
Model Construction Service

Cases
Case 1: Knock-out Cell Line Service
Experimental Objective
GenCRISPR™ technology was used to knock-out GS alleles in DG44 cells to construct stable cell lines. QC was used todetect the knock-out effect of target genes at DNA and functional levels.
Experimental Result
sgRNA cutting efficiency: The cutting efficiency of sgRNA designed by GenScript is greater than 80% (Fig.1)
Sequencing verification: The sequencing of selected monoclones proved that frameshift mutations occurred (Fig.2)
Functional testing: The target gene knock-out cells lost corresponding functions and could not grow in the medium

80
60
40
20

Wild-type CAAATAGGACCCTGTGAAGGAATCCGCATGGGAGATCATCTCTGGGTGGCC
GS-/CAAATAGGACCCTGTGAAGGAATCCGCATGGGAGATCATCT- -GGGTGGCC

0sgRNA1

sgRNA2

sgRNA3

Fig.1 sgRNA knock-out efficiency

Fig.2 Frameshift mutation occurred by knock-out GS gene monoclonal sequencing

3530
Viable cells number(×10^5 cell/ml)

Indel efficiency (%)

without glutamine (Fig.3)

25
0 mM-Glutamine
20

1 mM-Glutamine

15

2 mM-Glutamine
4 mM-Glutamine

10

8 mM-Glutamine

5
0
1

2

3

4

Day
Fig.3 Growth trend of DG44 cells with GS gene knock-out

34

Genome Editing
Model Construction Service

Case 2: Full-length gene knock-out cell line service
Experimental Objective
Conventional knockout induces gene knock-out by introducing frame-shift mutation, but in some special cases thetarget protein is still partially expressed and can be detected by some specific antibodies.
The vast majority of antibodies on the market have not been specifically verified, and many antibodies still detect thetarget protein bands in conventional knocked-out cells (Fig.4). The use of full-length knockout eliminates the concernof having target proteins remaining and shortens the research cycle.
M

1

2

3

4

5

120 KD--80 KD-Antibody from provider A
Fig.4 Knock-out cell line samples of supplier A for antibody detection(1-4 are samples after conventional knock-out of the target sequence, and 5 is wild-type control)

Experimental Result
Experiment 1. PCR and sequencing: PCR amplification was performed on two monoclones of genotype A*+B*(lane 1), genotype A (lane 2) and genotype B (lane 3) samples (primer design scheme is shown in Fig.5). The wholetarget gene fragments of the three samples were knocked out (Fig. 6A.B) and the residues were reconnected(Fig.6C). Sanger sequencing was performed to confirm the accuracy of the results.
A. Amplification
results of P1 sequence
M

P3-F

P1

P2

Target Gene
(Planned knock-out)

1

2

3

NC

B. Amplification
results of P2 sequence
M

1

2

3

NC

C. Amplification
results of P3 sequence
M

1

2

3

NC

P3-R
GenomeFig.5 Design scheme for verifying full-length geneknock-out effect by PCR amplification

Fig.6 Results for verifying full-length gene knock-out by PCR amplification(In A-C, lane 1: genotype A + B, lane 2: genotype A, lane 3: genotype B)*Genotype A does not contain screening labels, while genotype B contains
screening labels, both of which meet the delivery criteria

M NC 1

Experiment 2. Western Blot: The target protein marked by the red arrow was
knocked out. Since full-length knockout cell line service was used, it could bedetermined that the green arrow was not a target protein that is degraded or
incompletely expressed, but another protein with significantly increased
expression in the knock-out cell line (Fig.7). Therefore, the interference was
excluded.

35

2

3

Fig.7 Verification of full-length gene knockout
effect by Western Blot (1-3 are samples offull-length gene knock-out cells)

Genome Editing
Model Construction Service

Case 3: Knock-in Cell Line Service
Cell lines constructed by CRISPR gene knock-in were used for point mutation, over-expression and tracer genes
processes.
sgRNA+Cas9

Repair template

Mutated sequence or over-expressed gene or tracer gene

Experiment 1
Objective: Point mutation study
The sequence containing point mutation was knocked into the target site to construct cell lines by gene knock-in
technology, and the changes and mechanism caused by point mutation can be studied in physiological state.
Experimental Result
Sanger sequencing: Gene knock-in was used to complete point mutation, and successful point mutation wasverified by sequencing (Fig.8).
C G

Positive clone

Parental cell

Fig.8 Sequencing results of positive clone of point mutation

36

400

Objective: Over-expression study
300

The protein expression levels of 293 cell lines constructed by different
over-expression methods were compared.
Experimental Result
Protein expression levels: S2 and S3 were the protein expression levelsof cell line samples constructed by KI method, which were significantly
higher than that of S1 samples transfected by plasmids (Fig.9).

Titer(mg/L)

Genome Editing
Model Construction Service

S1 protein production in 293 cells

Experiment 2

200

100

0

S1

S2
S3
Stable cell poolFig.9 Protein expression levels of different
over-expression schemes (S1: plasmidtransfection, S2, S3: KI cell line)

Experiment 3
Objective: Tracer gene study
GenCRISPR™ technology is used to knock the target gene (GFP) into specific endogenous genes and construct fusion
proteins to trace specific endogenous genes.
Experimental Result
Flow cytometry (FACS): Significant GFP expression could be detected in positive clones by detecting the protein
expression in negative control cells, knock-in cell pools, and monoclones (Fig.10).

Parental cell

Cell pool

Positive clone

GFP

GFP

GFP

Fig.10 Expression of knock-in tracer gene GFP detected by FACS

37

Experimental Objective
CRISPR gene editing was performed on host cells through gRNA libraries targeting different loci, and different target
genes were screened out under different pressure conditions. Example: It is necessary to screen the genes that cause
host cells to be sensitive to 6-thioguanine. The wild type fails to grow with 6-thioguanine addition; after knock-out of
the target gene, it is resistant to 6-thioguanine, and it can grow normally with 6-thioguanine addition (Fig.12).

Cas9 viral particles

*
*
*

*

*

*

*

*

Pooled Library

*

*
*

*

*

*

Cas9 over-expressed Host*

Host cells

6-thioguanine
positive screen

gRNA viral library

Screened LibraryFig.11 Process of sorting target gene using gRNA libraryExperimental Result
Screening results: gRNA library was used for gene editing, 6-thioguanine was added for pressure sorting, and target
genes HPRT1 and NUDT5 were sorted after the comparison and evaluation of NGS sequencing (Fig.13).
After the target genes HPRT1 and NUDT5 were knocked out, the ratio of knock-out cells to total cells was increased
by adding 6-thioguanine (Fig.14), suggesting that the target genes were related to the sensitivity of 6-thioguanine,and host cells could grow in 6-thioguanine after knocking out.
Before treatment

After treatment-LogFDR

KO efficiency(%)

80
60
40
20
ND

0
Score
Fig.12 False discovery rate(FDR) and score of gRNA library sorting results(candidate genes marked)

HPRT1

NUDT5

Fig.13 Enrichment of gene knock-out cells under pressure

38

Genome Editing
Model Construction Service

Case 5: Sorting target loci using gRNA libraryGenome Editing
Model Construction Service

GenCRISPR™ Microbial Genome Modification
GenScript introduces microbial genome modification services for gene knock-in, knock-out and replacement ofbacterial (E.coli), saccharomyces cerevisiae, etc. For E.coli, GenScript has developed a novel λRed-CRISPR/Cas
technique that combines traditional λRed recombination with CRISPR/Cas9 to achieve traceless target gene editing.
Conventional Method

GenScript's New Service

NEW

Sorting Label

Target Gene

CRISPR cutting genome
FRT

FRT

Service Features
Traceless editing

Accurate to base

Easy sorting: No need for selective
labeling

Multiplex genome-editing: capable of knocking out up to
three genes simultaneously

Service Details
Service Items
E.coli gene knock-out service

E.coli gene knock-in andreplacement services

Provided by customers

Delivery standard

Delivery time

QC report on
glycerobacteriaof recombinant strain

At least 4-5 weeks

Strains and information to be modified
Name or sequence of a knock-out gene
Strains and information to be modified
Name or sequence of a gene to be knocked in*Insertion site/replacement sequence*GenScript can synthesize the gene sequence to be knocked in for you
We really appreciate the original plasmid of microbial genome modification provided by Prof. Yang Sheng and Dr. Jiang Yu of Shanghai Institutes for
Biological Sciences, Chinese Academy of Sciences (reference: Multigene editing in the Escherichia coli genome via the CRISPR-Cas9 system, Appl
Environ Microbiol 2015 Apr; 81 (7): 2506-2514).
This plasmid has been modified by GenScript's R&D team to meet the needs of microbial genome editing services.
*For GenCRISPR™ microbial genome modification service, please send an email to oligo@genscript.com or call 1-877-436-7274, and our professional
technical support team will be at your service.

39

04

Resource Center

Bioinformatics Tools............................................................ 41
Technical Guidance Materials......................................... 42
Articles Published by Customers.................................... 43
Customer Feedback.............................................................46

Resource Center

Bioinformatics Tools
CRISPR Design Tool for sgRNA & HDR-template
www.genscript.com/gencrispr-grna-design-tool.html
Enter the target Gene ID or Gene Symbol to quickly complete the gRNA & HDR-template design online. Based on thealgorithm developed and verified by published authoritative literatures, the evaluation parameters such as gRNA
localization, on-target /off-target score and comprehensive ranking are provided to help you select the appropriate
gRNA sequence.

Input interface

Output result

Genome-wide sgRNA Database
https://www.genscript.com/gRNA-database.html
An on-line genome-wide gRNA database for Human and Mouse can be applied to search for gRNA targeting any
gene and support the selection of downstream CRISPR gene editing or CRISPR transcriptional activation applications.

CRISPR gRNA Design Tool (enter sequence)
Change to https://www.genscript.com/tools/gRNA-design-tool
The target sequence is entered, the gRNA is designed
online for any customized sequence, and 12 commonlyused species are available. Galgorithms developed andvalidated by Zhang Feng Laboratory are applied tosupport your personalized research needs.

41

Resource Center

Technical Guidance Materials
From gene editing introduction and application to experimental details, GenScript provides you with comprehensive
knowledge and support in various forms, such as paper version, electronic version and lecture video, to help you
obtain valuable information more quickly and efficiently.

w w w. g e n s c r i p t . c o m . c n

CRISPR Technical Manual

Genome Editing Service Manual

CRISPR KO/KI Protocol

Principle and experimental processof gene editing technology
Downstream applicationand case analysis

Key points of CRISPR
experimental reagent selection
One-stop CRISPR customized
service offered by GenScript

HEK293T&293 / Jurkat / T cell
Thermo Fisher / Lonza / MaxCyte
Electroporation Platform

Case Study

CRISPR Webinar

Increase editing efficiency andreduce off-target rates in GCT &animal mode generation

Classical literature and real-time
analysis of hot topics
Concise interpretation of gene
editing knowledge

Welcome to visit GenScript website - Genome Editing Resource Center for the above information:(https://www.genscript.com/crispr-services.html#resources)
Or scan the QR code on the left

42

Resource Center

Articles Published by Customers
In terms of services and products, GenScript have been cited by more than 1,300 international famous academic
journals such as Cell, Science and PNAS. GenScript's gene editing services and products have been used by a numberof world-renowned institutions to publish their scientific achievements, demonstrating once again GenScript's ability tohelp scientists “Make Research Easy” in the industry.

The following are some scientific articles selected
Title: Genome-edited powdery mildew resistance in wheat without growth penalties
Journal: Nature IF: 43.07
Doi: 10.1038/s41586-022-04395-9
Title: High-yield genome engineering in primary cells using a hybrid ssDNA repair template and small-molecule
cocktails

Journal: Nat Biotechnol IF: 41.667
Doi: 10.1038/s41587-022-01418-8
Title: Re-engineering the adenine deaminase TadA-8e for efficient and specific CRISPR-based cytosine base editing
Journal: Nat Biotechnol IF: 41.667
Doi: 10.1038/s41587-022-01532-7
Title: A deletion mutation in TaHRC confers Fhb1 resistance to Fusarium head blight in wheat.
Journal: Nat Genet IF: 27.959
Doi: 10.1038/s41588-019-0425-8
Title: Transfer of cGAMP into bystander cells via LRRC8 volume-regulated anion channels augments STING-mediatedinterferon responses and anti-viral immunity

Journal: Immunity IF: 22.845
Doi: 10.1016/j.immuni.2020.03.016
Title: Myristoleic acid produced by enterococci reduces obesity through brown adipose tissue activation
Journal: Gut IF: 17.943
Doi: 10.1136/gutjnl-2019-319114
Title: Fast and sensitive detection of SARS-CoV-2 RNA using suboptimal protospacer adjacent motifs for Cas12a
Journal: Nat Biomed Eng IF: 17.135
Doi: 10.1038/s41551-022-00861-x
43

Resource Center

Title: The oncomicropeptide APPLE promotes hematopoietic malignancy by enhancing translation initiation
Journal: Mol Cell IF: 14.714
Doi: 10.1016/j.molcel.2021.08.033
Title: γ-6-Phosphogluconolactone, a Byproduct of the Oxidative Pentose Phosphate Pathway, Contributes to AMPK
Activation through Inhibition of PP2A.

Journal: Mol Cell IF: 14.714
Doi: 10.1016/j.molcel.2019.09.007
Title: High-Throughput and Efficient Intracellular Delivery Method via a Vibration-Assisted Nanoneedle/Microfluidic
Composite System

Journal: ACS Nano IF: 13.942
Doi: 10.1021/acsnano.2c07852
Title: Identification of human CD4+ T cell populations with distinct antitumor activity
Journal: SCIENCE ADVANCES IF: 12.804
Doi: 10.1126/sciadv.aba7443
Title: Programmable CRISPR-Cas9 microneedle patch for long-term capture and real-time monitoring of universal
cell-free DNA

Journal: Nat Commun IF: 12.124
Doi: 10.1038/s41467-022-31740-3
Title: SpG and SpRY variants expand the CRISPR toolbox for genome editing in zebrafish
Journal: Nat Commun IF: 12.124
Doi: 10.1038/s41467-022-31034-8
Title: Lupus enhancer risk variant causes dysregulation of IRF8 through cooperative lncRNA and DNA methylation
machinery

Journal: Nat Commun IF: 12.124
Doi: 10.1038/s41467-022-29514-y
Title: VviPLATZ1 is a major factor that controls female flower morphology determination in grapevine
Journal: Nat Commun IF: 12.124
Doi: 10.1038/s41467-021-27259-8
Title: Aspartate aminotransferase Rv3722c governs aspartate-dependent nitrogen metabolism in Mycobacterium
tuberculosis

Journal: Nature Communications IF: 11.878
Doi: 10.1038/s41467-020-15876-8
44Resource Center

Title: dbGuide: a database of functionally validated guide RNAs for genome editing in human and mouse cells
Journal: Nucleic Acids Research IF: 11.147
Doi: 10.1093/nar/gkaa848
Title: A programmable omnipotent Argonaute nuclease from mesophilic bacteria Kurthia massiliensis
Journal: Nucleic Acids Res IF: 10.162
Doi: 10.1093/nar/gkaa1278
Title: Effector gene silencing mediated by histone methylation underpins host adaptation in an oomycete plant
pathogen.

Journal: Nucleic Acids Res IF: 10.162
Doi: 10.1093/nar/gkz1160
In terms of services and products, GenScript have been cited nearly 10,000 times by more than 1300 journals of
biomedicine such as Cell, Nature, Science and PNAS.
Please visit the customer's published literature athttps://www.genscript.com.cn/reference_peer-reviewed_literature.html.

10000

Molecular Biology Services

9000
8000

Catalog Products

7000

Peptide Services

6000

Protein Services

5000

Antibody Services

4000

Oligo Services

3000

Bioinformatics Tools

2000

Gene Editing Services

1000
0

COVID192012 2013 2014 2015 2016 2017 2018 2019 2020 2021 2022

Number of Literatures in GenScript from2010 to 2022

Proportion Distribution of GenScript Products
and Services in the Literature

The growth of GenScript is inseparable from the support of our customers. Our value is fully reflected by the publicationof papers and academic achievements. In order to thank our customers for their continuous love to GenScript, and more
importantly, to thank the front-line researchers for their contribution to the progress of life science for all mankind,
GenScript has launched an award-winning activity for publishing articles.

45Resource Center

Customer Feedback"GenScript's CRISPR service was a life saver for my research. We were totally stuck on a specific point
mutation in our E.coli strain, and GenScript was able to step in and get it done. Updates are provided
through every step of the process and they've been a pleasure to deal with."
Chad Johnston, Ph.D., Banting Postdoctoral Fellow in the Collins Lab at MIT.

"Long ssDNA sequences are difficult to produce in the lab, especially at the high concentrations
necessary for gene editing experiments, we were able to successfully integrate large DNA sequences into
primary human T cells using GenScript's long ssDNA product."Dr. Theodore Roth from the Marson Lab at University of California San Francisco.

46

05

Ordering Guide
and Contact Information

Order Method..........................................................................48
Order Query............................................................................. 48

Ordering Guide and Contact Information

Order Method
GenScript website
CRISPR/Cas9 Service

On-line Ordering
Enter the Ordering
Information

Submit
Add to Shopping
Cart

Close an AccountComplete theOrdering

On-line Ordering:Log in on-line ordering system, fill in and submit the ordering information, add to yourshopping cart, close an account, and finally complete the ordering. SgRNA, ssDNA andCRISPR plasmids can be ordered on-line.

Ordering by
Customer Service:

Submit the inquiry form containing order information (can be downloaded at the bottom of
the Service page on the official website of GenScript) and send it to oligo@genscript.com, andour professional technical support team will be at your service.

Order Query
How to query?

Log on GenScript website

1. Log in to your GenScript account
2. Click Account Name - User Center

User Center

3. Click "My Order/Query" in the taskbar on the left of the page
4. Select "All Types of Orders” in the Order TypeMy Order/Query

5. Click the order number to enter the "Order Details" page to view theorder progress. For delayed or difficult orders, please email us for
consultation and confirmation. We will reply and follow up as soon as
possible.

Select Service

Click the Order Number

For delayed or difficult orders, please email us for consultation andconfirmation. We will reply and follow up as soon as possible.

48

Memo No.
Date

Memo No.
Date

GenScript is committed to supporting customer needs andto bringing cutting-edge technologies to labs globally

Explore more service & application information!
www.genscript.com
1-877-436-7274
gene@genscript.com
860 Centennial Ave. Piscataway NJ 08854