CRISPR Gene Editing
GMP Cas9 Nucleases
Overview
Ultra-performance with aseptic manufacturing, traceable documentation, and stringent testing.
크리스퍼 유전자 편집 관련 응용 분야가 기초 연구에서 임상 제조 분야로 발전함에 따라 고품질 원료와 완벽한 문서화가 더욱 중요해지고 있습니다.
점점 더 엄격해지는 사양을 충족하기 위해 GenScript는 GMP 등급 sgRNA 합성 서비스와 함께 GMP 등급 GenCRISPR™ Ultra SpCas9 및 eSpCas9 Nucleases를 제공합니다.
GMP 등급 GenCRISPR ™ Ultra SpCas9 및 eSpCas9 Nucleases는 무균 제조, 추적 가능한 문서화 및 엄격한 테스트를 포함하되 이에 국한되지 않는 관련 우수 제조 관행 (GMP) 지침에 따라 제조됩니다.
GMP 등급 Ultra SpCas9 및 eSpCas9 Nucleases는 테스트된 세포주 및 일차 T 세포에서 높은 knockout and knock-in 효율로 크리스퍼 유전자 교정을 가능하게 합니다.
Highlights
| Consistent High Editing Efficiency | High Lot-to-lot Consistency |
| • 세포주 및 1차 T 세포에서 테스트 • 다양한 electroporation scale • 대부분의 Ultra Cas9 Nucleases 사용 |
•In in-vitro cleavage assay •In gene editing applications |
| Ultra-High Stability | Stringent Specifications |
| • -25°C 및 4°C에서 가속 테스트 수행 • -20°C에서 실시간 안정성 테스트 수행 • 동결-해동 안정성 테스트 수행 |
•무균 제조 •추적 가능한 제조공정 •엄격한 QC 테스트 |
Product List
GMP Grade SpCas9 and eSpCas9
| Cat. No. | Product Name |
| Z03623-GMP | GenCRISPR™ Ultra NLS-Cas9-GMP |
| Z03624-GMP | GenCRISPR™ Ultra eSpCas9-2NLS-GMP |
Basic GMP Grade SpCas9 and eSpCas9
| Cat. No. | Product Name |
| Z03623 | GenCRISPR™ Ultra NLS-Cas9-basic GMP |
| Z03624 | GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP |
RUO Grade SpCas9 and eSpCas9
| Cat. No. | Product Name |
| Z03621 | GenCRISPR™ Ultra NLS-Cas9-Research |
| Z03622 | GenCRISPR™ Ultra eSpCas9-2NLS-Research |
Stringent Manufacturing and Release Specifications
Manufacturing Specifications
| Test | GenCRISPR™ Ultra Cas9 and eSpCas9 Nucleases | ||
| GMP Grade* | Basic GMP Grade** | RUO Grade | |
| Facility | GMP Compliant | Non-GMP Compliant | Non-GMP Compliant |
| cGMP Guidelines | +++ | ++ | + |
| ISO9001 | / | / | +++ |
| ISO13485 | +++ | +++ | / |
| Quality Control | +++ | +++ | + |
| Documentation | +++ | ++ | + |
*GMP Grade는 GenScript가 엄격한 공정 제어와 완전한 문서 기록을 통해 현재 우수 제조 관리 기준(cGMP), ISO 13485 품질 관리 시스템 표준의 지침을 준수하여 GMP 준수 시설에서 제조된 Cas9 nucleases를 설명하기 위해 사용하는 특정 용어입니다. GenScript는 특정 지역에서 프로젝트를 적용하는 데 도움이 되는 문서, 현장 감사 및 기타 지원을 제공할 수 있습니다.
**Basic GMP Grade는 ISO 13485 품질 관리 시스템 표준을 준수하고 보다 엄격한 공정 제어와 비교적 완전한 문서 기록을 통해 제조된 시약을 설명하기 위해 GenScript가 사용하는 브랜드 용어입니다. GenScript는 특정 지역에서 프로젝트를 적용하는 데 도움이 되는 문서, 현장 감사 및 기타 지원을 제공할 수 있습니다.
Release Specifications
| Test | GenCRISPR™ Ultra Cas9 and eSpCas9 Nucleases | |||
| GMP Grade | Basic GMP Grade | RUO Grade | ||
| Concentration | 10.0 mg/mL±10% | 10.0 mg/mL±10% | 10.0 mg/mL±10% | |
| Appearance | Clear, colorless | Clear, colorless | Clear, colorless | |
| Purity | SDS-PAGE | ≥ 95 % | ≥ 95 % | ≥ 90 % |
| SEC-HPLC | ≥ 95 % | ≥ 95 % | ≥ 95 % | |
| Activity | ≥ 90 % (SpCas9) ≥ 85 % (eSpCas9) |
≥ 90 % (SpCas9) ≥ 85 % (eSpCas9) |
≥ 90 % (SpCas9) ≥ 85 % (eSpCas9) |
|
| Endotoxin | < 10 EU/mg | < 10 EU/mg | < 10 EU/mg | |
| Residual DNase | ≤ 10 ng/mg | ≤ 10 ng/mg | Non-detectable Qualitative Method |
|
| Residual RNase | ≤ 1 ng/mg | ≤ 1 ng/mg | Non-detectable Qualitative Method |
|
| Residual HCP | < 10 ng/mg | < 10 ng/mg | ||
| Residual HCDNA | ≤ 1 ng/mg | ≤ 1 ng/mg | ||
| Mycoplasma | < LOD | < LOD | ||
| Sterility | Sterile | Sterile | ||
Case Study
Consistent high knockout efficiency in primary T cells using different grade Cas9 nucleases
Figure 1: TRAC knock-out in primary T cells using GMP, basic GMP and RUO Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 with electroporation |
Figure 1: TRAC knock-out in primary T cells using GMP, basic GMP and RUO Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 with electroporation The cells were transfected with GenCRISPR™ Ultra NLS-SpCas9-Research (Cat. No. Z03621), GenCRISPR™ Ultra NLS-SpCas9-basic GMP (Cat. No. Z03623), GenCRISPR™ Ultra NLS-SpCas9-GMP (Cat. No. Z03623-GMP) or GenCRISPR™ Ultra eSpCas9-2NLS-Research (Cat. No. Z03622), GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP (Cat. No. Z03624), GenCRISPR™ Ultra eSpCas9-2NLS-GMP (Cat. No. Z03624-GMP) and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services ) for human TRAC gene knock-out by electroporation. After transfection and cell culture, the TRAC knock-out efficiency was measured by FACS. This data indicated that GMP, basic GMP and RUO Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 enable consistent high gene editing efficiency in primary T cells. |
| Cell Type: | Primary T cells |
| Transfection Method: | Electroporation |
| Target Gene: | TRAC |
| Cell amount: | 1.0 x 106 cells |
| Cas9: sgRNA ratio: | 1:2 |
| RNP amount: | 25 pmol |
Consistent high editing efficiency in primary T cells in different electroporation scale
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| Figure 2: TRAC knock-out in primary T cells using GMP Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 with electroporation in different electroporation scale
Human primary T cells were cultured for the test. 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 Cas9 Nucleases from other suppliers, and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services) for human TRAC gene knock-out by electroporation in a small electroporation scale of 20 μl and a large scale of 100 μl. After transfection and cell culture, the TRAC knock-out efficiency was measured and analyzed by FACS. The data indicated that GMP Grade GenCRISPR™ Ultra Cas9 and GenCRISPR™ Ultra eSpCas9 maintain consistent high editing efficiency in different electroporation scales |
| Electroporation Scale | 20 μl | 100 μl |
| Cas9 nuclease amount | 4 μg | 20 μg |
| Cell amount | 1.0 × 106 | 5.0 × 106 |
| Cas9: sgRNA (molar ratio) | 1:2 | 1:2 |
| RNP amount | 25 pmol | 125 pmol |
High knock-in efficiency in primary T cells using GMP Grade GenCRISPR™ Ultra Cas9 and Ultra eSpCas9
Figure 3: 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 measured and 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: | Electroporation |
| Target Gene: | TRAC |
| Gene Knocked-in: | GFP |
| Cell amount: | 1.0 x 106 cells |
| Cas9: sgRNA ratio: | 1:2 |
| RNP amount: | 25 pmol |
High knockout efficiency in cell lines using GMP Grade GenCRISPR™ Ultra Cas9 and Ultra eSpCas9
Figure 4: CIITA, GM-CSF, TGFBR2 knock-out in Jurkat cells using GenCRISPR™ Ultra SpCas9 or GenCRISPR™ Ultra eSpCas9 with electroporation |
Jurkat cells were cultured for the test. The cells were transfected with GenCRISPR™ Ultra NLS-SpCas9-GMP (Z03623-GMP) or GenCRISPR™ Ultra eSpCas9-2NLS-GMP (Z03624-GMP) or wild type Cas9 protein from other supplier and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services) for human CIITA, GM-CSF and TGFBR2 gene knock-out by electroporation. After transfection and cell culture, the gene editing efficiency was analyzed by Sanger sequencing. Both GenScript GenCRISPR™ Ultra NLS-SpCas9-GMP (Z03623-GMP) and GenCRISPR™ Ultra eSpCas9-2NLS-GMP (Z03624-GMP) show high gene knock-out efficiency. |
| Cell Type: | Jurkat cells |
| Transfection Method: | Electroporation |
| Target Gene: | CIITA, GM-CSF and TGFBR2 |
| Cell amount: | 0.5 x 106 cells |
| Cas9: sgRNA ratio: | 1:3 |
| RNP amount: | 7.5 pmol |
Resources
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CRISPR RNP User Manual.pdf | A guide on how to use GenScript’s RNP products for targeted genome editing |
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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 |
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GenCRISPR gRNA Design Tool.pdf | Interactive CRISPR guide RNA design tool |
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GenCRISPR™ Gene Editing Solutions.pdf | All solutions you need for a successful CRISPR gene editing project |
제품 및 서비스 문의
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