GenCRISPR™ Ultra eSpCas9-2NLS-GMP
GenCRISPR™ Ultra ESpCas9-2NLS-GMP

Figure 2: TRAC knock-out in primary T cells using GMP Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 in different electroporation scale.

GenCRISPR™ Ultra ESpCas9-2NLS-GMP

Figure 4: CIITA, GM-CSF and TGFBR2 knockout in Jurkat cells using GMP Grade GenCRISPR™ Ultra SpCas9 or GenCRISPR™ Ultra eSpCas9 Nuclease.

GenCRISPR™ Ultra ESpCas9-2NLS-GMP

Figure 3: GFP gene knock-in at TRAC site in primary T cells using GMP Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 Nuclease.

GenCRISPR™ Ultra ESpCas9-2NLS-GMP

Figure 1: TRAC knock-out in primary T cells using GMP, basic GMP and RUO Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 Nuclease.

GenCRISPR™ Ultra eSpCas9-2NLS-GMP

GenCRISPR™ Ultra eSpCas9-2NLS-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 two nuclear localization signals (NLS) expressed with the Cas9 nuclease, the RNP complex enters the nucleus and cleaves 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.
Z03624-GMP
¥1,250,632.00

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Overview
Description GenCRISPR™ Ultra eSpCas9-2NLS-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 two nuclear localization signals (NLS) expressed with the Cas9 nuclease, the RNP complex enters the nucleus and cleaves 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.

Nucleases 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.

Properties
Source Recombinant mutant Cas9 with nuclear localization signal (NLS) at both N-terminal and C-terminal 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
Application sgRNA-dependent double-stranded DNA cleavage.
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. 

Quality Control Specifications
Appearance Clear, colorless, liquid
Identity Consistent with the theoretical sequence by LC-MS/MS
Purity ≥ 95.0 as analyzed by SDS-PAGE
≥ 95.0 as analyzed by SEC-HPLC
Concentration by A280 10.0 mg/ml ± 1.0 mg/ml
Bioactivity (in vitro) ≥ 85.0%
Residual DNase ≤ 10 ng/mg
Residual RNase ≤ 1 ng/mg
Endotoxin Level < 10 EU/mg
Residual HCP ≤ 10 ng/mg
Residual HCD ≤ 1 ng/mg
Mycoplasma Non-detectable by Culture method
Sterility No growth

Examples
  • GenCRISPR™ Ultra ESpCas9-2NLS-GMP
    • GenCRISPR™ Ultra ESpCas9-2NLS-GMP

    Figure 2: TRAC knock-out in primary T cells using GMP Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 in different electroporation scale.

  • GenCRISPR™ Ultra ESpCas9-2NLS-GMP
    • GenCRISPR™ Ultra ESpCas9-2NLS-GMP

    Figure 4: CIITA, GM-CSF and TGFBR2 knockout in Jurkat cells using GMP Grade GenCRISPR™ Ultra SpCas9 or GenCRISPR™ Ultra eSpCas9 Nuclease.

  • GenCRISPR™ Ultra ESpCas9-2NLS-GMP
    • GenCRISPR™ Ultra ESpCas9-2NLS-GMP

    Figure 3: GFP gene knock-in at TRAC site in primary T cells using GMP Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 Nuclease.

  • GenCRISPR™ Ultra ESpCas9-2NLS-GMP
    • GenCRISPR™ Ultra ESpCas9-2NLS-GMP

    Figure 1: TRAC knock-out in primary T cells using GMP, basic GMP and RUO Grade GenCRISPR™ Ultra Cas9 or GenCRISPR™ Ultra eSpCas9 Nuclease.


Background
References
  1. 1. Jinek, Martin, et al. "A programmable dual-RNA–guided DNA endonuclease in adaptive bacterial immunity." science 337.6096 (2012): 816-821. 
  2. 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. 3. Larson, Matthew H., et al. "CRISPR interference (CRISPRi) for sequence-specific control of gene expression." Nature protocols 8.11 (2013): 2180-2196.
  4. 4. Ran, F. Ann, et al. "Genome engineering using the CRISPR-Cas9 system." Nature protocols 8.11 (2013): 2281-2308. 
  5. 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.