| Description |
GenCRISPR? Ultra NLS-Cas9-GMP is utilized for
CRISPR gene editing applications. The Cas9 nuclease forms a stable
ribonucleoprotein (RNP) complex with the guide RNA (gRNA) component. With the
help of nuclear localization signal (NLS) expressed with the Cas9 nuclease, the
RNP complex enters the nucleus and cleaves the target gene. When compared with a
plasmid-based delivery system, the RNP delivery system has been observed to
increase the on-target gene editing efficiency and decrease off-target effects. |
| Selection Guide |
GenScript provides both wild-type and mutant
Cas9 nucleases in GMP Grade* for selection based on your specific downstream
applications. The GMP Grade Cas9 nucleases can be utilized as ancillary
materials for Cell, Gene, and Tissue-Based Products and are manufactured under
the standards listed below:
- § USP <1043>. Ancillary
materials for Cell, Gene and Tissue-engineered products.
- § ICH Q7 Good Manufacturing Practice Guide for Active Pharmaceutical
Ingredients
- § NMPA: Technical guidelines for pharmaceutical research and
evaluation of immune cell therapy products
- § Guideline and procedure specified in Chinese Pharmacopeia.
|
Nuclease
|
Description
|
GenCRISPR? Ultra
NLS-Cas9-GMP
(Cat. No. Z03623-GMP)
|
Recombinant
Cas9 nuclease (wild-type) of Streptococcus
pyogenes; ideal choice for most CRISPR gene editing applications where
high editing efficiency is preferred.
|
|
GenCRISPR? Ultra
eSpCas9-2NLS-GMP
(Cat.
No. Z03624-GMP)
|
Recombinant
enhanced specificity Cas9 nuclease (mutant) of Streptococcus pyogenes; ideal
for CRISPR gene editing applications which require low off-target effects and
robust on-target cleavage.
|
|
|
sgRNA-dependent double-stranded DNA cleavage. |
| Source |
Recombinant Cas9 nuclease (wt) with an N-terminal NLS expressed in
E.coli |
| Species |
S. pyogenes |
| Tag |
Tag-free
|
| Theoretical Molecular Weight |
160 kDa
|
| Concentration |
10.0 mg/ml
|
| Active temperature |
Optimal at 37 °C.
|
| Storage Buffer |
25 mM Tris-HCl, 300 mM NaCl, 0.1 mM EDTA, 50% Glycerol, pH 8.0. |
| Storage & Stability |
Store at -20±5°C for up to 12 months from the date of manufacture. Avoid repeated freeze-thaw cycles. |
| Appearance |
Clear, colorless, liquid
|
| Purity |
≥ 95.0% as analyzed by SDS-PAGE
≥ 95.0% as analyzed by SEC-HPLC |
| Concentration |
10.0 mg/ml ± 1.0 mg/ml |
| Bioactivity |
≥ 90.0% |
| Residual DNase |
≤ 10 ng/mg
|
| Residual RNase |
≤ 1 ng/mg
|
| Endotoxin Level |
< 10 EU/mg
|
| Host Cell Protein |
≤ 10 ng/mg |
| Host Cell DNA |
≤ 1 ng/mg |
| Mycoplasma |
< LOD
|
| Sterility |
No growth |
Figure 1: TRAC knock-out in primary T cells using GMP, basic GMP and RUO Grade GenCRISPR? Ultra Cas9 or GenCRISPR? Ultra eSpCas9 Nuclease.
Figure 2: TRAC knock-out in primary T cells using GMP Grade GenCRISPR? Ultra Cas9 or GenCRISPR? Ultra eSpCas9 in different electroporation scale.
Figure 4: CIITA, GM-CSF and TGFBR2 knockout in Jurkat cells using GMP Grade GenCRISPR? Ultra SpCas9 or GenCRISPR? Ultra eSpCas9 Nuclease.
Figure 3: GFP gene knock-in at TRAC site in primary T cells using GMP Grade GenCRISPR? Ultra Cas9 or GenCRISPR? Ultra eSpCas9 Nuclease.
| References |
- 1. Jinek,
Martin, et al. "A programmable dual-RNA–guided DNA endonuclease in
adaptive bacterial immunity." science 337.6096 (2012):
816-821.
- 2. Chen,
Sean, et al. "Highly efficient mouse genome editing by CRISPR
ribonucleoprotein electroporation of zygotes." Journal of
Biological Chemistry 291.28 (2016): 14457-14467.
- 3. Larson,
Matthew H., et al. "CRISPR interference (CRISPRi) for sequence-specific
control of gene expression." Nature protocols 8.11
(2013): 2180-2196.
- 4. Ran, F.
Ann, et al. "Genome engineering using the CRISPR-Cas9 system." Nature
protocols 8.11 (2013): 2281-2308.
- 5. Kim,
Sojung, et al. "Highly efficient RNA-guided genome editing in human cells
via delivery of purified Cas9 ribonucleoproteins." Genome research 24.6
(2014): 1012-1019.
|
| Annotation |
*GMP
Grade is a specific term that GenScript uses to describe the Cas9 nucleases
manufactured in GMP-complaint facility and in compliance with guidelines of
Current Good Manufacturing Practice (cGMP), ISO 9001 and ISO 13485 quality
management system standards with stringent process controls and complete
documentation records. GenScript is capable of providing documents, site
audits, and other support to help with the applications of your projects in
specific regions. |
For laboratory research or clinical research and manufacturing of cell-based products. Direct human use, including taking orally and injection are forbidden.
