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Genome-CRISP™ CRISPR-Cas9 stable cell lines

Introduction

GeneCopoeia's stable cell lines constitutively expressing the CRISPR Cas9 nuclease enable you to carry out CRISPR genome editing applications with high efficiency.

The Genome-CRISP™ Cas9 stable cell lines are available pre-made in many human cell lines, and in mouse cell line Neuro2a. The CRISPR Cas9 nuclease is stably integrated into either the human AAVS1 or mouse ROSA26 “Safe Harbor” sites (Figure 1), or randomly via lentiviral-mediated transduction.

The CRISPR Cas9 nuclease, in combination with single guide RNAs (sgRNAs), is widely used to create targeted genomic modifications, from gene knockouts, mutagenesis, fusion tagging, and more, in eukaryotic cells and animal models. Stable Cas9 integration ensures robust and consistent expression of the CRISPR Cas9 nuclease.  

Advantages

  • Stable Cas9 integration minimizes the need for co-transfection or co-transduction of sgRNAs, ideal for high-throughput sgRNA applications.
  • Single clone isolation provides consistent, high-level Cas9 expression in a uniform genetic background.
  • Cas9 functionally validated using the IndelCheck™ T7 Endonuclease I assay (Figure 2).
  • Compatible with GeneCopoeia Genome-CRISP™ sgRNA clones,sgRNA libraries,and donor clones.

 

Figure 1. Structure of a stably integrated CRISPR Cas9 nuclease construct. Hygromycin (Hygro) selection is used for isolation of single clones. CBh: Hybrid CBA promoter. 3X FLAG: N-terminal 3X FLAG tag. NLS: nuclear localization signal. bGH pA: Bovine growth hormone polyadenylation signal. EF1a: Elongation factor 1a promoter. copGFP: Copepod green fluorescent protein. T2A: Element to allow translation of copGFP and Puro independently from the same transcript. SV40 pA: Simian virus 40 polyadenylation signal.

 

Cas9 expression from within these stable cell lines exhibits high activity, as shown by the T7 Endonuclease I mismatch cleavage assay (Figure 2).

Cas9 stable cell lline T7EI assay validation

 

 

Figure 2. Demonstration of stable Cas9 activity in a cell line. Human H1299 cells stably expressing Cas9 nuclease from the AAVS1 Safe Harbor site were transfected with a plasmid carrying an sgRNA for the HUWE I gene. 24 hours post-transfection, cells were harvested and used for genomic DNA extraction. Using the GeneCopoeia IndelCheck™ kit, PCR products flanking the sgRNA target site were generated, denatured, reannealed, and treated with (+) or without (-) T7 Endonuclease I, which cleaves mismatched DNA. The presence of two bands below the uncut band at the correct sizes indicates that Cas9 is active in these cells.

 

To Order

Order pre-made Cas9-expressing stable cell lines

Choose your human or mouse Cas9-expressing stable cell line from the list below. On this page, you can also purchase Safe Harbor Cas9 knock-in kits, for do-it-yourself engineering of the Cas9-expressing stable cell line of your choice. In addition, you can purchase one of our IndelCheck™ insertion/deletion detection kits, for efficient validation of CRISPR sgRNAs and TALENs, and for screening for CRISPR- or TALEN-modified cell lines

Buy Catalog # Cell line Description Selection marker Cas9 integration site Price, Academic Price, Industry
Human Cas9-expressing stable cell lines
SL501 H1299 Human cell line H1299 stably expressing CRISPR Cas9, single clone Puro AAVS1 $1395 $2695
SL502 HEK293T Human cell line HEK293T stably expressing CRISPR Cas9, single clone Puro AAVS1 $945 $1995
SL503 HeLa Human cell line HeLa stably expressing CRISPR Cas9, single clone Hygro AAVS1 $1395 $2695
SL504 A549 Human cell line A549 stably expressing CRISPR Cas9, single clone Hygro AAVS1 $1395 $2695
SL514 MCF-7 Human cell line MCF-7 stably expressing CRISPR Cas9, single clone Hygro AAVS1 $1395 $2695
SL515 MDA-MB-231 Human cell line MDA-MB-231 stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL516 MDA-MB-468 Human cell line MDA-MB-468 stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL517 T47D Human cell line T47D stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL518 HepG2 Human cell line HepG2 stably expressing CRISPR Cas9, single clone Puro AAVS1 $1395 $2695
SL520 AGS Human cell line AGS stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL521 BXPC-3 Human cell line BXPC-3 stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL522 SNU-475 Human cell line SNU-475 stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL523 HT-29 Human cell line HT-29 stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL525 LoVo Human cell line LoVo stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL527 KATO111 Human cell line KATO111 stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL535 SK-BR-3 Human cell line SK-BR-3 stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
SL544 HCC70 Human cell line HCC70 stably expressing CRISPR Cas9, single clone Hygro Random $1395 $2695
Mouse Cas9-expressing stable cell lines
SL509 Neuro2a Mouse cell line Neuro2a stably expressing CRISPR Cas9, single clone Hygro ROSA26 $1395 $2695
SL510 Neuro2a Mouse cell line Neuro2a stably expressing CRISPR Cas9, single clone Puro ROSA26 $1395 $2695
SL511 Neuro2a Mouse cell line Neuro2a stably expressing CRISPR Cas9, single clone Neo ROSA26 $1395 $2695

  MSDS

 

Note: GeneCopoeia also offers custom-built Cas9-expressing stable cell line services. To inquire, please contact inquiry@genecopoeia.com.

 

Human AAVS1 and mouse Rosa26 Safe Harbor Cas9 knock-in kits

Don't see the Cas9-expressing stable cell line you're looking for? You can choose from a series of human AAVS1 and mouse ROSA26 Safe Harbor knock-in kits (below), each with Cas9-expressing donor clone. These products enable you to conveniently engineer your cell line of interest to express the CRISPR-Cas9 nuclease.

The human AAVS1 and mouse ROSA26 Safe Harbor Cas9 knock-in kits are part of GeneCopoeia's system of Safe Harbor knock-in kits and ORF clones. For more information, visit GeneCopoeia's human AAVS1 and mouse ROSA26 Safe Harbor products page.

Buy Catalog # Product Description Price Cas9 clone User manual
Human AAVS1 Safe Harbor Cas9 knock-in kits
SH012 Genome-CRISP™ Cas9 human AAVS1 Safe Harbor knockin kit-Puro CRISPR-based kit for knock-in of Cas9 nuclease to human AAVS1 Safe Harbor site-puromycin selection. Includes CRISPR-Cas9 clone targeting AAVS1, Cas9-AAVS1 donor clone with puromycin selection, and verification PCR primer pairs. $1595 DC-C9NU-03
SH013 Genome-CRISP™ Cas9 human AAVS1 Safe Harbor knockin kit-Hygro CRISPR-based kit for knock-in of Cas9 nuclease to human AAVS1 Safe Harbor site-hygromycin selection. Includes CRISPR-Cas9 clone targeting AAVS1, Cas9-AAVS1 donor clone with hygromycin selection, and verification PCR primer pairs. $1595 DC-C9NU-04
SH014 Genome-CRISP™ Cas9 human AAVS1 Safe Harbor knockin kit-Neo CRISPR-based kit for knock-in of Cas9 nuclease to human AAVS1 Safe Harbor site-G418 selection. Includes CRISPR-Cas9 clone targeting AAVS1, Cas9-AAVS1 donor clone with G418 selection, and verification PCR primer pairs. $1595 DC-C9NU-05
Mouse ROSA26 Safe Harbor Cas9 knock-in kits
SH062 Genome-CRISP™ Cas9 mouse ROSA26 Safe Harbor knockin kit-Puro CRISPR-based kit for knockin of Cas9 nuclease to Mouse ROSA26 Safe Harbor site-puromycin selection. Includes CRISPR-Cas9 clone targeting ROSA26, Cas9-ROSA26 donor clone with puromycin selection, and verification PCR primer pairs $1595 DC-C9NU-06
SH063 Genome-CRISP™ Cas9 mouse ROSA26 Safe Harbor knockin kit-Hygro CRISPR-based kit for knockin of Cas9 nuclease to Mouse ROSA26 Safe Harbor site-hygromycin selection. Includes CRISPR-Cas9 clone targeting ROSA26, Cas9-ROSA26 donor clone with hygromycin selection, and verification PCR primer pairs $1595 DC-C9NU-07
SH064 Genome-CRISP™ Cas9 mouse ROSA26 Safe Harbor knockin kit-Neo CRISPR-based kit for knockin of Cas9 nuclease to Mouse ROSA26 Safe Harbor site-G418 selection. Includes CRISPR-Cas9 clone targeting ROSA26, Cas9-ROSA26 donor clone with G418 selection, and verification PCR primer pairs $1595 DC-C9NU-08

 

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CRISPR/TALEN Insertion or Deletion Detection System

GeneCopoeia's IndelCheck™ CRISPR/TALEN Insertion or deletion detection system is the perfect companion to Cas9-expressing stable cell lines. The system allows you to readily 1) Validate/estimate the cleavage efficiency of target site-specific CRISPR sgRNAs and TALENs in cell culture; and 2) Screen mammalian cell clones for CRISPR- or TALEN-mediated modifications.

For more information, visit GeneCopoeia's IndeCheckTM CRISPR/TALEN insertion or deletion detection system products page

Buy Catalog # Product Description Price
IC001 IndelCheck™ CRISPR/TALEN insertion or deletion detection system (50 rxns) Includes target site PCR kit (IC003) and T7 endonuclease I assay kit (IC005) $206
IC002 IndelCheck™ CRISPR/TALEN insertion or deletion detection system (200 rxns) Includes target site PCR kit (IC004) and T7 endonuclease I assay kit (IC006) $622
IC003 Target site PCR Kit (version 2.0) , 50 rxns* PCR reagents for amplifying region flanking CRISPR/TALEN target site, prior to T7 Endonuclease I digestion* $159
IC004 Target site PCR Kit (version 2.0), 200 rxns* PCR reagents for amplifying region flanking CRISPR/TALEN target site, prior to T7 Endonuclease I digestion* $479
IC005 T7 endonuclease I assay kit, 50 rxns Cleave mismatched PCR products using T7 endonuclease I to detect indel mutations $99
IC006 T7 endonuclease I assay kit, 200 rxns Cleave mismatched PCR products using T7 endonuclease I to detect indel mutations $299
  IC007 Target site PCR cloning kit, 20 rxns Clone PCR products for sequencing and identification of CRISPR- or TALEN-mediated genomic modifications Coming soon
  IC008 Target site PCR cloning kit, 100 rxns Clone PCR products for sequencing and identification of CRISPR- or TALEN-mediated genomic modifications Coming soon

*Version 2.0 now conains buffer, SuperHeRo™ DNA polymerase, and nucleotides together in one mix for greater convenience compared with Version 1.0.

   

 

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Questions? Please contact us at inquiry@genecopoeia.com or 1-866-360-9531.

Applications

Applications

  • High-throughput knockout screening with many sgRNAs, either individually or in pools. Ideal for drug target discovery (Figure 3).
  • Convenient validation of several drug target candidates.
  • Validation of sgRNA cleavage activity in a fast-growing, easy-to-transfect or transduce model cell line, either prior to transfection/transduction of your cell line, or to troubleshoot sgRNAs with little or no cleavage activity. 
  • Appications requiring inducible genome modification.

 

Figure 3. Example application of Cas9-expressing stable cell lines Use of the Genome-CRISP™ Cas9 stable cell lines with CRISPR sgRNA libraries for drug target discovery. A. Pooled screen. Cas9-expressing cells infected with each sgRNA library pool are screened for the desired readout. Pooled cells are subjected to Sanger sequencing for individual sgRNAs, or deep sequencing to look for over- or under-representation of individual sgRNAs. B. Knockout screen using arrayed sgRNAs. Cas9-expressing cells are infected with individual sgRNA lentiviruses. Wells are screened for the readout of interest. Individual sgRNAs corresponding to the phenotype of interest are already known without sequencing.