Cas9 / Cas12a / Cas13a Nucleases for precise RNA-guided DNA or RNA editing and detection

CRISPR 유전자 편집 기술은 유전자 및 세포 치료, 세포주 엔지니어링, 동물 모델 생성, 체외 진단을 포함한 다양한 다운스트림 애플리케이션에서 활용되고 있습니다.
GenScript는 MIT 및 하버드의 유수의 연구실과의 파트너십을 통해 검증된 GMP 등급 및 RUO 등급의 Cas9, Cas12a and Cas13a nuclease, sgRNA 합성 및 HDR donor template 합성서비스 등을 제공하고 있습니다.

SpCas9 Nuclease (wild type)
Recombinant wild-type S. pyogenes Cas9 nucleases, purified from E. coli expression and contains various nuclear localization sequences (NLSs).

eSpCas9 Nuclease (mutant)
Recombinant mutant S. pyogenes Cas9 nucleases, purified from E. coli expression and contain various nuclear localization sequences (NLSs).

Cas9-eGFP Nuclease
Recombinant S. pyogenes Cas9 nuclease and eGFP fusion protein, purified from E. coli expression, and contains nuclear localization sequence (NLS) and C-terminal 6xHis tag.

SaCas9 Nuclease
Recombinant S. aureus Cas9 nuclease, purified from E. coli expression and contains nuclear localization signals (NLSs) and no tag.

Cas9 Nickase D10A
Recombinant S. pyogenes Cas9 Nickase D10A, purified from an E. coli strain expressing the protein, and contains nuclear localization sequences (NLSs).

Cas12a Nuclease
Recombinant Acidaminococcus sp. (strain BV3L6) Cas12a (cpf1) nuclease or Lachnospiraceae bacterium ND2006 Cas12a nuclease, purified from E. coli expression, and contains nuclear localization sequences (NLSs).

Cas13a Nuclease
Recombinant Leptotrichia wadei Cas13a nuclease or Leptotrichia buccalis Cas13a nuclease, purified from E. coli expression.

Gene Editing Kits

Cas Nuclease Antibodies

High knockout efficiency in primary T cells (Cat. No. Z03623 and Z03624)

Figure 1:TRAC knockout in primary T cells using basic GMP Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 with electroporation

The cells were transfected with GenCRISPR™ Ultra NLS-SpCas9-basic GMP (Cat. No. Z03623) or GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP (Cat. No. Z03624), and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services) for human TRAC gene knockout by electroporation. After transfection and cell culture, the TRAC knock-out efficiency was analyzed by FACS. This data indicated that both basic GMP Grade GenCRISPR™ Ultra Cas9, and GenCRISPR™ Ultra eSpCas9 enable consistent and high gene knockout efficiency in primary T cells. Note: No Cargo: No Cas9 protein added (Negative control) No EP: No electroporation for cell transfection (Negative control) D3: KO efficiency evaluated after day 3 D6: KO efficiency evaluated after day 6

High knock-in efficiency in primary T cells (Cat. No. Z03623-GMP and Z03624-GMP)

Figure 2: GFP gene knock-in at TRAC site in primary T cells using GMP Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 with electroporation

The cells were transfected with GMP Grade GenCRISPR™ Ultra NLS-SpCas9-GMP (Cat. No. Z03623-GMP) or GenCRISPR™ Ultra eSpCas9-2NLS-GMP (Cat. No. Z03624-GMP), and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services), as well as dsDNA HDR template (synthesized by GenScript HDR Donor Template Synthesis Services) by electroporation. After transfection and cell culture, the gene knock-in efficiency was analyzed by FACS. Both GenScript GenCRISPR™ Ultra NLS-SpCas9-GMP (Cat. No. Z03623-GMP), and GenCRISPR™ Ultra eSpCas9-2NLS-GMP (Cat. No. Z03624-GMP) maintain high gene knock-in efficiency in primary T cells.

High knock-in efficiency in primary T cells (Cat. No. Z03623-GMP and Z03624-GMP)

Figure 3: GFP Gene knock-in at TRAC loci in primary T cells by using GenCRISPR™ Cas9 v1.1 or GenCRISPR™ Cas9 v1.2 and sgRNA with electroporation.

The cells were transfected with GenCRISPR™ Cas9 v1.1 (Cat. No. Z03701) or GenCRISPR™ Cas9 v1.2 (Cat. No. Z03702), and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services), as well as dsDNA HDR template (synthesized by GenScript HDR Donor Template Synthesis Services) by electroporation. After transfection and cell culture, the gene knock-in efficiency was analyzed by FACS. Both GenCRISPR™ Cas9 v1.1 and GenCRISPR™ Cas9 v1.2 with optimized NLSs enable high gene knock-in efficiency.

High knockout efficiency in Jurkat cells (Cat. No. Z03623-GMP and Z03624-GMP)

Figure 4: CIITA, GM-CSF and TGFBR2 Gene Knock-out in Jurkat cells by using GMP Grade GenCRISPR™ Ultra NLS-Cas9-GMP or Ultra eSpCas9-2NLS GMP with electroporation

The cells were transfected with GenCRISPR™ Ultra NLS-SpCas9-GMP (Cat. No. Z03623-GMP), or GenCRISPR™ Ultra eSpCas9-2NLS-GMP (Cat. No. Z03624-GMP), or a competitor’s product, and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services) for gene knockout by electroporation. After transfection and cell culture, the knockout efficiency was analyzed by Sanger sequencing. This data shows that both GMP Grade GenCRISPR™ Ultra NLS-Cas9, and GenCRISPR™ Ultra eSpCas9-2NLS enable high gene knockout efficiency in Jurkat cells.

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High knockout efficiency in NK-92 cells (Cat. No. Z03623)

Figure 5: CD96 and NKG2A gene knockout in NK-92 cells using GenCRISPR™ Ultra NLS-Cas9-basic GMP with electroporation.

NK-92 cells were cultured for the test. The cells were transfected with GenCRISPR™ Ultra NLS-Cas9-basic GMP (Cat. No. Z03623) and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services) for CD96 or NKG2A gene knock-out by electroporation. After transfection and cell culture, the gene editing efficiency was analyzed by Sanger sequencing. The results show that GenCRISPR™ Ultra NLS-Cas9-basic GMP (Z03623) has high gene knockout efficiency in NK-92 cells for both genes tested.

Gene Editing with low off-target effects in 293T cells (Cat. No. Z03623 and Z03624)

Figure 5: TRAC gene knockout in 293T cells by using GenCRIPSR™ Ultra NLS-Cas9-basic GMP or GenCRIPSR™ Ultra eSpCas9-2NLS-basic GMP and sgRNA with electroporation. A) TRAC gene knockout efficiency analysis. B) Off-target effects analysis on OT1 site. C) Off-target effects analysis on OT2 site. The cells were transfected separately with either GenCRIPSR™ Ultra NLS-Cas9-basic GMP, or GenCRIPSR™ Ultra eSpCas9-2NLS-basic GMP, or a Competitor’s product, and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services) for TRAC gene knockout by electroporation. OT1 and OT2 were predicted as the off-target sites and were selected for off-target effects analysis. After transfection and cell culture, the knockout efficiency and off-target effects were analyzed by Sanger sequencing. This data indicates that basic GMP Grade GenCRISPR™ Ultra eSpCas9-2NLS engaged gene editing has low off-target effects, as well as robust on-target editing efficiency.

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