Edit-R™ Gene Editing Workflows
Choose the right tools for your application
Whether you’re goal is a functional protein knockout for your gene of interest,
running a loss-of-function screen, or creating an insertion or other knock-in, thisworkflow guide will assist you in selecting the right Edit-R™ gene editing reagentsfor your application.

Choose your application
Gene knockout
Gene knock-in

Loss-of-function
screening of multiple
genes at onceSingle gene knockout

precise insertion or alteration of a geneKnockout cell line creation
or loss-of-function analysis
in cell population

AAAACas9

Transfect Edit-R Cas9 mRNA orprotein NLS for transient expression
with no risk of DNA integration andfewer off-targets.

Cas9
Cas9
Use Edit-R Lentiviral Cas9
nuclease for stable orinducible cell line creation for
optimal editing efficiency.

Chooseguide RNA

Transduce
lentiviralsgRNA pools

Synthetic guide RNAs work best fortransient activity, and required nocloning or purification!

Optimize Cas9
 elivery a ndd xpression
eAND guideRNA delivery

Cas9 + guide RNA transfected cells

3 daysAssess g eneeditingefficiencyandfunctional
knockout
phenotype

3 daysCreating aknockout cell line?
Single colony expansion
in 96-well plates.

Loss-of-function
analysis in cell
population.

2-3 weeksMC

1Choose aCas9 reagent
and createstable c ell line

PM

C1

P

Perform
pooled screen
experiment

AnalyzeenrichedsgRNA
constructs inReference vs.
Experimental
sampleCas9 cell line mixed
population
Cas9 cell line mixed
population

Choose lentiviral particles
for the creation of stable orinducible Cas9 cell lines withchoice of optimal promoter
for your cell type.

Choose aCas9 r eagentand create
stabecell line
Transduce cells at
MOI < 1sgRNA

Edit-R lentiviral sgRNA pools
are fully sequenced libraries
of algorithm-designed
sgRNA demonstrating efficientgene editing at single-copy
integrations.
Collect Reference sample, apply selective
pressure to Experimental sample
Reference sample

Choose guide
RNA anddeliver to cells

Cas9

Use Edit-R Lentiviral Cas9
nuclease for stable orinducible cell line creation for
optimal editing efficiency.

Experimental sample

Selection withblasticidin and cellline expansion

Edit-R synthetic guide RNA isavailable in arrayed predefined genecollections or custom libraries

Isolate gDNA

Experimental gDNA

Large insert

or
Short insert

Synthetic guide RNAs work best fortransient activity, and required nocloning or purification!

Optimize Cas9
delivery andexpression ANDguide RNA as
well a s donor
oligo delivery

DNA donor
plasmid for
insertionof >50 nt
Single-stranded
DNA oligo donor
for insertionsof <50 nt

Enrichment using
FACS or antibiotic resistance(when applicable)
Cas9 + custom guide RNA + donor
DNA mixed population

Expansion of selected cells

3 daysPerformarrayed screen
experiment

Assess 
loss-of-function
phenotype

Single colonyexpansion into
96-well plates

Delivery with appropriate
DharmaFECT transfection
reagent formulation

3 daysReference gDNA

Choose guide
RNA anddonor oligo
sourceExperimental sample

Ready to learn more about these offerings?
Contact your local Horizon Discovery representative or
email us at ts.dharmacon@horizondiscovery.com

Transfect Edit-R Cas9 mRNA orprotein NLS for transient expression
with no risk of DNA integration andfewer off-targets.

MC1

Assess HDRefficiency
Detection of HDR
modification/insertion RFLP orjunctional PCR

Assess loss-of-function
with a phenotypic assay

Characterizeclonal cell line

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2-3 weeks

MC1P

V20621

Choose a
Cas9 reagent

Choose a
Cas9 r eagent

Arrayed screening forone-gene-per-well analysis

Pooled screening with noneed for automation

AAAACas9