CRISPR Gene Editing
Cas9 / Cas12a / Cas13a Nucleases
Overview
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.
Highlights
RNP-mediated CRISPR Gene Editing | Consistent high knockout and knock-in efficiency | High Lot-to-lot Consistency |
• 비바이러스성, 유전자 통합 위험 없음 • 빠른 편집 작용 및 분해 • 낮은 세포 독성 및 면역원성 |
• 세포주 및 1차 T 세포에서 테스트 • 다양한 electroporation scale • 대부분의 Ultra Cas9 Nucleases 사용 |
• In in-vitro cleavage assay • In gene editing applications |
Selection Guide
Applications | Recommendations | Cat. No. | Product Names |
Gene KO/KI with high editing efficiency | Ultra SpCas9 (wt) | Z03621 | GenCRISPR™ Ultra NLS-Cas9-Research |
Z03623 | GenCRISPR™ Ultra NLS-Cas9- basic GMP | ||
Z03623-GMP | GenCRISPR™ Ultra NLS-Cas9-GMP | ||
Gene KO/KI with low off-target effects | Ultra eSpCas9 (mt) | Z03622 | GenCRISPR™ Ultra eSpCas9-2NLS-Research |
Z03624 | GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP | ||
Z03624-GMP | GenCRISPR™ Ultra eSpCas9-2NLS-GMP | ||
Gene KI with high KI efficiency | SpCas9 with optimized NLS | Z03701 | GenCRISPR™ Cas9 v1.1 |
Z03702 | GenCRISPR™ Cas9 v1.2 | ||
Gene KO/KI following with flow cytometry or other fluorescent assays | SpCas9-eGFP fusion protein | Z03393 | GenCrispr NLS-Cas9-EGFP Nuclease |
Z03467 | |||
Gene KO/KI with lower MW Cas nucleases | AsCas12a | Z03502 | GenCRISPR™ Cas12a (Cpf1) Nuclease |
LbCas12a | Z03753 | GenCRISPR™ LbCas12a Nuclease | |
SaCas9 | Z03699 | GenCRISPR™ SaCas9 2NLS Nuclease | |
in vitro Diagnostics | LwaCas13a | Z03486 | GenCRISPR™ Cas13a (C2c2) Nuclease |
LbuCas13a | Z03742 | GenCRISPR™ LbuCas13a Nuclease | |
AsCas12a | Z03502 | GenCRISPR™ Cas12a (Cpf1) Nuclease | |
LbCas12a | Z03753 | GenCRISPR™ LbCas12a Nuclease |
Product List
SpCas9 Nuclease (wild type)
Recombinant wild-type S. pyogenes Cas9 nucleases, purified from E. coli expression and contains various nuclear localization sequences (NLSs).
Cat. No. | Product Name |
Z03385 | GenCrispr Cas9-C-NLS Nuclease |
Z03386 | GenCrispr Cas9 Nuclease |
Z03388 | GenCrispr Cas9-N-NLS Nuclease |
Z03389 | GenCrispr NLS-Cas9-NLS Nuclease |
Z03468 | GenCrispr Cas9-N-NLS Nuclease |
Z03469 | GenCrispr NLS-Cas9-NLS Nuclease |
Z03621 | GenCRISPR™ Ultra NLS-Cas9-Research |
Z03623 | GenCRISPR™ Ultra NLS-Cas9-basic GMP |
Z03623-GMP | GenCRISPR™ Ultra NLS-Cas9-GMP |
Z03701 | GenCRISPR™ Cas9 v1.1 |
Z03702 | GenCRISPR™ Cas9 v1.2 |
eSpCas9 Nuclease (mutant)
Recombinant mutant S. pyogenes Cas9 nucleases, purified from E. coli expression and contain various nuclear localization sequences (NLSs).
Cat. No. | Product Name |
Z03470 | GenCrispr eSpCas9-N-NLS Nuclease |
Z03622 | GenCRISPR™ Ultra eSpCas9-2NLS-Research |
Z03624 | GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP |
Z03624-GMP | GenCRISPR™ Ultra eSpCas9-2NLS-GMP |
Z03692 | GenCRISPR™ Ultra eSpCas9-N-NLS Research, tag-free |
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.
Cat. No. | Product Name |
Z03393 | GenCrispr NLS-Cas9-EGFP Nuclease |
Z03467 | GenCrispr NLS-Cas9-EGFP Nuclease |
SaCas9 Nuclease
Recombinant S. aureus Cas9 nuclease, purified from E. coli expression and contains nuclear localization signals (NLSs) and no tag.
Cat. No. | Product Name |
Z03699 | GenCRISPR™ SaCas9 2NLS Nuclease |
Cas9 Nickase D10A
Recombinant S. pyogenes Cas9 Nickase D10A, purified from an E. coli strain expressing the protein, and contains nuclear localization sequences (NLSs).
Cat. No. | Product Name |
Z03390 | GenCrispr NLS-Cas9-D10A Nickase |
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).
Cat. No. | Product Name |
Z03502 | GenCRISPR™ Cas12a (Cpf1) Nuclease |
Z03753 | GenCRISPR™ LbCas12a Nuclease |
Cas13a Nuclease
Recombinant Leptotrichia wadei Cas13a nuclease or Leptotrichia buccalis Cas13a nuclease, purified from E. coli expression.
Cat. No. | Product Name |
Z03486 | GenCRISPR™ Cas13a (C2c2) Nuclease |
Z03742 | GenCRISPR™ LbuCas13a Nuclease |
Gene Editing Kits
Cat. No. | Product Name |
L00688 | GenCrispr Mutation Detection Kit |
L00689 | GenCrispr sgRNA Screening Kit |
L00694 | GenCrispr sgRNA Synthesis Kit |
Z03396 | GenCrispr T7 Endonuclease I |
Cas Nuclease Antibodies
Cat. No. | Product Name |
A01885 | GenCrispr Cas9 Antibody, pAb, Rabbit |
A01935 | GenCRISPR™ SpCas9 Antibody (4A1), mAb, Mouse |
A01936 | GenCRISPR™ SpCas9 Antibody (14B6), mAb, Mouse |
A01951 | GenCRISPR™ SaCas9 Antibody (11C12), mAb, Mouse |
A01952 | GenCRISPR™ SaCas9 Antibody (26H10), mAb, Mouse |
Case Study
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) |
Cell Type: | Primary T cells |
Transfection Method: | Lonza 4D-Nucleofector® Transfection System |
Target Gene: | TRAC |
Cell amount: | 1.0 x 106 cells |
Cas9: sgRNA ratio: | 1:2 (25 pmol : 50 pmol) |
Cas9 Nucleases: | Z03623 GenCRISPR™ Ultra NLS-Cas9-basic GMP Z03624 GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP |
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. |
Cell Type: | Primary T cells |
Transfection Method: | Lonza 4D-Nucleofector® Transfection System |
Target Gene: | TRAC |
Cell amount: | 1.0 x 106 cells |
Cas9: sgRNA ratio: | 1:2 (25 pmol : 50 pmol) |
HDR donor: | 2 μg |
Cas9 Nucleases: | Z03623 GenCRISPR™ Ultra NLS-Cas9-basic GMP Z03624 GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP |
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. |
Cell Type: | Primary T cells |
Transfection Method: | Lonza 4D-Nucleofector® Transfection System |
Target Gene: | TRAC |
Gene Knocked-in: | GFP |
Cell amount: | 1.0 x 106 cells |
Cas9: sgRNA ratio: | 1:2 (25 pmol : 50 pmol) |
Cas9 Nucleases: | Z03623 GenCRISPR™ Ultra NLS-Cas9-basic GMP Z03624 GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP |
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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Cell Type: | Jurkat cells |
Transfection Method: | Neon™ electroporation system |
Target Gene: | CIITA, GM-CSF, TGFBR2 |
Cell amount: | 0.5 x 106 cells |
Cas9: sgRNA ratio: | 1:3 (7.5 pmol: 22.5 pmol) |
Cas9 Nucleases: | Z03623 GenCRISPR™ Ultra NLS-Cas9-basic GMP Z03624 GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP |
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. |
Cell Type: | NK-92 cells |
Transfection Method: | Lonza 4D-Nucleofector® Transfection System |
Target Gene: | CD96 , NKG2A |
Cell amount: | 0.4 x 106 cells |
Cas9: sgRNA ratio: | 1:1.2 |
RNP amount: | 80 pmol |
Cas9 Nucleases: | Z03623 GenCRISPR™ Ultra NLS-Cas9-basic GMP Z03624 GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP |
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. |
Cell Type: | 293T cells |
Transfection Method: | Neon™ Electroporation Transfection System |
Target Gene: | TRAC |
Cell amount: | 0.2 x 106 cells |
Cas9: sgRNA ratio: | 1:3 |
Cas9 Nucleases: | Z03623 GenCRISPR™ Ultra NLS-Cas9-basic GMP Z03624 GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP |
Resources
CRISPR RNP User Manual.pdf | A guide on how to use GenScript’s RNP products for targeted genome editing | |
CRISPR RNP Handbook.pdf | Enabling RNP-mediated CRISPR gene editing and transforming life science research | |
Cas9 and sgRNA Protocol.pdf | Simple protocol for CRISPR gene editing using GenCRISPR™ Cas9 nuclease | |
GenCRISPR gRNA Design Tool.pdf | Interactive CRISPR guide RNA design tool | |
GenCRISPR™ Gene Editing Solutions.pdf | All solutions you need for a successful CRISPR gene editing project |
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