Thank You...

Kroma! iD

Direct Definitive Genomics

For more information:

Christopher Tompkins
President and GM
ctompkins @kromatid.com
303-819-5246


Structural Error Free Cells: 72% 14

Correct Edit
Site Structure

. : ,
we Correct Edit ©
Site Structure
Two SVs ~ ©
Correct Edit
Site Structure
One SV Fey > Structural
Error Free ©

Cells

KromatiD


Two dGH Assay Designs 5

De Novo dGH Targeted dGH

Strand Specific

Strand Specific
Chromatid PAIN

Targeted PROBES Edit _—— Inversion

=
—

Signal on the “Dark” Chromosome
indicates a change and orientation and
thus and Inversion

Va s from
f Ti ) Reference Genome
Structure

Variations from
Expected Target
Structure


Application Use Cases for Gene Editing

Material Qualification

RM
Prod

»

dGH

Kb

Reject

y

Subject Assessment

Kroma

f)

dGH

} Edit

Optimization

Edit

»

dGH

SV SV
Rate Spec

Process Changes |

Quality Control

Edit

»

dGH

Kh

Reject

y



De Novo dGH 10

Translocated Proprietary Strand Specific
Inversian Fragment Chromatid Paints measure
structural variations from the
reference genome

© Inversion
Translocation

eo Source

’ Inversion

Inversion

#


dGH Integrates with Editing Workflow

Edit } Expand } Edited Cell Population
>
DNA
Sample b Prep b Pooled DNA Analysis
Orthogonal
> ae
ne Single Stranded Me hi
ingle Stranded Metaphase
Prep Chromosomes
ne —=
ES
— Strand Specific
§ Hyb Hybridization Probes 6
oo
oO
Any Fluorescence Imaging

2 Image System
i=
a —r

Analyze Image Scoring

Kroma



dGH Integrates with Editing Workflow

Edit } Expand } Edited Cell Population
>
DNA .
Sample b Prep } Pooled DNA Analysis
> ae

Kroma


Next Steps— Whole Genome dGH 2.0

dGH 2.0 —

High Throughput Imaging

Cell
Prep

f= We

\ a

Kroma

Whole Genome
Coverage

Al Image

Analysis

100%

90
80
70
8 60

I 0

.

z 40

z 30
20

Structure Class and Type

Routine efficient measurement of
comprehensive structural genomic
profiles

Qualification
Optimization
Screening
Control

NIH National Human Genome Research Institute Fast -Track SBIR


Classes of Structural Errors

100%
90
80
70
2
& 60
8 50 ONLY Eait Site MIXED Edit Site and NO Edit Site
a involvement Random/Homologous Involvement
8 40 Site Involvement
J
o
5 30
oa
* § 5 2
20 5 8 — 2 a
, 8 x 8 8 x 8 &§
a 2 g rs 2 ¢ 2 <8 re
a o -
10 E £ E 5 s—2 E : s
8 E
—— ee oes ee
Class 0: Error Class 1: Edit Site Class 2: Mixed Class 3: Other
Free Cells Structural Errors Structural Errors Structural Errors

Kro m a “Possible edit site structures depend on number and location of edits


Kroma

dGH Analysis Tools for Gene Editing

Single Cell Structural Genomic Analysis


Targeted dGH 12

Proprietary Strand Specific

Correct Edit Chromatid Probes measure
Site Structure variations from expected edit
structure

= Edit Site
Edit Site Dual Edit L_ s © Inversion
Inversion

bad Correct Edit
Site Structure

HEK-293FT Cells


Structural Variation Rates

Mouse C1-C4

©

Inversion

TiD

Inversion

©)

11


Structural Error Free Cells

100%

90

80

70

)

oO

© 60

= 50

Ww

re}

2& 40

[~4

oa

5 30

a

a ® a

Ss a 5 g

2 g 5 52 3 a

° s 2 8 5s 8 § » § &§&
a ¢ $ 2 2 5 = 2

10 - § é 3 s 8 e #2 3
5 FO Be 8 fF &
——— ee oes

Class 0: Error
Free Cells

Kro m a *Possible edit site structures depend on number and location of edits


Key Characteristics of dGH

Directional Genomic Hybridization

| Direct measurement of structural

variations in many single cells

* Complete characterization of SVs possible
including inversions

¢ Retains the full cellular and chromosomal
context without pooling DNA

* Definitive detection from image data without
bioinformatic interpretation

¢ High resolution method with LODs of 4 KB and

De Novo dGH <0.5% prevalence

Kroma


Application Use Cases for Gene Editing

Material Qualification

=>

Reject

Kroma


Application Use Cases for Gene Editing

Material Qualification

RM
Prod

»

dGH

Kb

Reject

y

Subject Assessment

Kroma

f)

dGH

} Edit

Optimization

Edit

»

dGH

Di.

SV
Rate

/

SV
Spec

Process Changes |



Complex Mis-Repair 3

Edit Site Homologous Site Random
DSB DSB DSB

Low Frequency

. Process Dependent
High Frequency

Low Frequency

Genome Dependent

Opportunities for Mis-Repair are Proportional to the number
of Double Strand Breaks. Rates of DSBs likely depend on
the Edit System, Raw Materials and Conditions

TiD


Our Roots...

Human Lymphocytes

EFFECT--SVs Per Cell

DOSE—Radiation Gray

Deep experience measuring DNA mis-repair for
Dosimetry and Oncology
Kroma


Multiple Structural Variation

Correct Edit
Site Structure ©

Translocated
Fragment
Edit Site
Inversion

Edit Site
Translocation

©

TiD

13


Application Use Cases for Gene Editing

Material Qualification

RM
Prod

»

dGH

Reject

Kb

y

Kroma

Optimization

Edit

»

dGH

p Lim /

Process Changes |

SV
Spec