Gentaur Solutions

CRISPR Solutions – Reagents

CRISPR Solutions – KO Service

Cell Engineering Solutions

1. Recombinant
2. DNA Technology
3. Viral Vectors:
4. Lentivirus, Adenovirus
5. Cell Engineering
6. Techniques
7. Gene Editing
8. Gene Expression Studies
9. Protein Production
10. Disease Modeling
11. Gene Therapy
12. Vaccine Development
13. Regenerative Medicine

Clones

1. CRISPR ORF
2. ORF
3. miRNA
4. siRNA
5. CRISPR siRNA circRNA
6. miRNA circRNA
7. ORF
8. miRNA siRNA

Virusses

1. Lentivirus
2. Adeno-Associated Virus
3. Adenovirus
4. Retrovirus
5. HSV
6. Baculovirus

Characteristic Lentivirus Adenovirus AAV Retrovirus Packaging Capacity (Insert size)

9 kb 8.5 kb 4.5 kb 8 kb

Transduction Efficiency

Infection Most Dividing/

Non-Dividing Cells

Most Dividing/ Non[1]Dividing Cells (with high

transduction rate

towards primary cells)

All Cell Types

(depending on

Serotype)

Dividing Cells

Expression Stable Transient Transient or Stable Stable

Integrating Yes No Site-specific integration Yes

Immune Response

Genetic Material RNA Double Stranded Linear

DNA

Single Stranded Linear

DNA

RNA

● Low ●●●●● High

Viral Packaging

Lentivirus: 107~1010

Adenovirus: 1010~1012

AAV: 1010~1013

Retrovirus: 107

Gene Synthesis

Site-directed Mutagenesis

Subcloning

Fusion

Inducible Vectors

DNA Amplification

Start at any point …. ... or let us do it all

Gene Editing &

Cloning Gene Delivery

Promoters

Reporters & Tags

CMV • EF1a •

UbC •PGK •

H1 • U6 • CAGGS • MSCV

GFP • RFP • HA tag •

His tag • Luciferase •

Resistance Markers •

N-Terminal/C-Terminal

Reference

Lumbar Neuronal Cells after 4 weeks of

Intrathecal injection of AAV9-EGFP

Data Courtesy of Dr. Douglas Lopes, King's College, London

In vivo Application

To establish methods for intrathecal injection with

AAV9-GFP, then express desired transgene

Quantitative Western Analysis following infection with 3pfu

Null or DENND1a V2 adenovirus

Data from McAllister, J. et al PNAS, 111(15) pp. E1519-27.

In vitro Application

To test the role of DENND1A.V2 in promoting a PCOS

phenotype in human theca cells

Gene Editing Validation & Cloning

Gene Delivery Cell

Engineering

Supporting Products up- and downstream

Titer Kits

qPCR-based kit

for lentivirus, AAV,

or retrovirus

Transduction

Enhancers

Non-toxic formulation

to boost AAV, or

lentivirus efficiency

Ready-to-use

Models

Primary

Immortalized &

Stable Cell Lines

Expression

Assays

Extraction Kits

RT, PCR, qPCR

Custom Gene Synthesis & Cloning

Custom Vector Construction

Viral Packaging

DNA Amplification

By Product

ORF

CRISPR/Cas9 (KO, Activation)

miRNA & 3’UTR

siRNA/shRNA/RNAi

circRNA

Specialized Vectors: Cre-Inducible,

Organelle Labeling, mir-Tuners

Reference

Jennifer Doudna

University of California, Berkeley

Emmanuelle Charpentier

Max Planck Institute for Infection Biology

Nobel Prize in 2020: "Type V Restriction Enzyme"

Genomi

c DNA

Cas9

sgRNA

Cas9

Cas9 Proteins

CRISPR/Cas9 Vector/Virus Libraries

Cas9 Expressing Cell Lines

CRISPR Cloning Vectors

CRISPR Screening Kits & Enzymes

Comprehensive CRISPR Solutions

CRISPR

Base Editors

ABE (A→T)

CBE (C→G)

Comprehensive One-Stop Solution Provider

- Breadth of Products

- Our Genetic & Cell Line expertise

Personalized Solution for your Unique Needs

Cas9 & sgRNA Validation

Design

Gene Delivery Cell

Cas9 & sgRNA

Design

Gene Regulations with Cas9

Cas9 Variants

SpCas9

SaCas9

Cas12a (Cpf1)

Cas9 Types

Nuclease

Nickase

dCas9 with Effector

Array of Delivery Method

Gene Delivery

Characteristic Lentivirus AAV Retrovirus

Packaging Capacity

(Insert size)

9 kb 4.5 kb 8 kb

Transduction Efficiency

Infection Most Dividing/

Non-Dividing

Cells

All Cell Types

(depending on

Serotype)

Dividing Cells

Expression Stable Transient or

Stable

Stable

Integrating Yes Site-specific

integration

Yes

Immune Response

Genetic Material RNA Single Stranded

Linear DNA

RNA

23

Head-start with Cas9 Expressing Cell Lines

Cell

Engineering

Examples of Cas9 Ready Tumor Cells

T24 HCT116 A549 Jurkat

PC-3 HT1080 MDA-MB[1]468

A375

USO2 HuTu 80 MCF-7 Calu-6

HEL HK-2 A2780 SKUT-1

K562 293 HEY RL95-2

U-87 293T OVCAR3 CT26

HeLa 786-O INS1E NIH3T3

HT-29 Hep3B L3.6pl TM4

HCT-15 HepG2 THP-1 RCS

Screening and Validation made Easy

Validation

Surveyor Assay (T7E1)

Can only detect

presence of editing

Detect the nature of

edits

Cas9 Cleavage Assay

Screening and Validation made Easy

Validation

Surveyor Assay (T7E1)

Cas9 Cleavage Assay

depending on type of edit

o Hetero-Duplex (Monoallelic

Mutation or Heterozygous

Biallelic Mutation) = CUT

o Homo-duplex (Wild Type or

Homozygous Biallelic Mutation) =

UNCUT

It does NOT differentiate between monoallelic

mutation and heterozygous biallelic mutation.

Cas9 Cleavage Assay

Cas9 Cleavage Assay differentitates

between monallelic mutations and biallelic

mutations.

Viral Packaging

Lentivirus: 107~1010

Adenovirus: 1010~1012

AAV: 1010~1013

Retrovirus: 107

Gene Knockout

Gene Activation

Epigenetic Gene Repression

Epigenetic Gene Activation

Start at any point …. ... or let us do it all

sgRNA Cloning

Vector Design

Reporters

& Tags

Single • Multiplex

All-in-One • SgRNA

Only

GFP • RFP • HA tag •

His tag • Luciferase •

Resistance Markers •

N-Terminal/C-Terminal

RNP

Virus

Plasmid

What expression

type?

ie. Stable, transient

4

What insert

size?

Your gene of

interest,

reporter, marker

Let's Start a Conversation!

28

The abm Difference: 3 Partners for the Journey

Cas9 & sgRNA Validation

Design

Gene Delivery Cell

Engineering

Reducing Work:

We design the

sgRNA sequences

for your GOI using our

software.

Backstopping Uncertainty:

We provide another set of

3 sequences if preliminary

results are not effective

Mitigating Risks:

We provide guidance

on cell line generation,

tips on choosing

delivery method & drug

selection.

Decreasing Unknown:

We provide support in

deciphering screening

results and/or provide

screening services.

By Service

sgRNA Design & Vector Construction

Cell Line Engineering (CRISPR KO

Service)

Cas9 Proteins

CRISPR/Cas9 Vector/Virus Libraries

Cas9 Expressing Cell Lines

CRISPR Cloning Vectors

CRISPR Screening Kits & Enzymes

Other Reagents (e.g. Antibody)

Gentaur has also PCR Enzymes:

Cell Biology Products

Engineered for research or demanding diagnostic product applications, our

premium PCR, qPCR, RT-qPCR, and RT-PCR product development has culminated in

our world-class, specially formulated collection of enzymes, MasterMixes and Kits.

With years of expertise in the cell biology industry, abm scientists aim to deliver the

highest quality and most complete collection of cell culture products and services

bringing you the newest technologies at prices you can afford.

Category Product Name Cat. No. Size

Reverse Transcriptase

1. & RT-PCR All-In-One 5X RT MasterMix G592 100 rxn
2. qPCR Mastermixes BlasTaq™ 2X qPCR MasterMix G891 500 rxn
3. qPCR Virus Titer Kits
4. qPCR Lentivirus Titer Kit LV900 100 rxn
5. qPCR AAV Titer Kit G931 100 rxn
6. SafeView™ DNA Stains
7. Safe-Red™ Gel G680 1.0 ml
8. SafeView™ Classic G108 1.0 ml

Molecular Biology

1. Enzymes and Kits
2. RNase R E049 50 μl
3. Screen It™ CRISPR Cas9 Cleavage Detection Kit G990 100 Reactions
4. Product Name Cat. No. Size
5. DNAfectin™ Plus Transfection Reagent G2500 1.0 ml
6. Applied Cell Extracellular Matrix G422 25ml
7. MycoAway™ Mycoplasma Elimination Cocktail (1000X) G398 1.0 ml
8. Mycoplasma PCR Detection Kit G238 100 rxn
9. Mycoplasma Pro PCR Detection Kit G239 100 rxn