GMP Cas9 Nucleases

Ultra performance with aseptic manufacturing, traceable documentation, and stringent testing.

Overview Highlights Products Case Study Resources

Catalog Products » GenCRISPR™ Cas9 Genome Editing Products » GenCRISPR™ Ultra Nucleases

Overview

With CRISPR gene editing involved applications moving from basic research to clinical manufacturing, high-quality raw materials and complete documentation are becoming more essential. To fulfill the increasingly stringent specifications, GenScript offers GMP Grade GenCRISPR™ Ultra SpCas9 and eSpCas9 Nucleases, alongside GMP Grade sgRNA synthesis services.

GMP Grade GenCRISPR™ Ultra SpCas9 and eSpCas9 Nucleases are manufactured under the relevant guidelines for Good Manufacturing Practice (GMP), including but not limited to aseptic manufacturing, traceable documentation, and stringent testing. GMP Grade Ultra SpCas9 and eSpCas9 Nucleases enable CRISPR gene editing with high knockout and knock-in efficiency in tested cell lines and primary T cells.

Highlights

Consistent High Editing Efficiency

Consistent high knockout and knock-in efficiency

In tested cell lines and primary T cells
At different electroporation scale
Using different grade of Ultra Cas9 Nucleases

High Lot-to-lot Consistency

High lot-to-lot consistency

In in vitro cleavage assay
In gene editing applications

Ultra High Stability

Activity and purity maintain high and stable

In accelerated testing at 25°C and 4°C
In real-time stability testing at -20°C
In freeze-thaw stability testing

Stringent Specifications

Manufactured under guidelines for GMP, including

Aseptic manufacturing
Traceable documentation
Stringent testing

CRISPR Gene Editing

Product List

All

GMP Grade [11]

RUO Grade [11]

GMP Grade SpCas9 and eSpCas9

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RUO Grade SpCas9 and eSpCas9

Cat. No.	Product Name	Quantity	Price	Order
Z03622	GenCRISPR™ Ultra eSpCas9-2NLS-Research		$765.00

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	+++	++	+

Note:
*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 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.

**Basic GMP Grade is a branding term that GenScript uses to describe reagents manufactured in compliance with ISO 13485 quality management system standards and with more stringent process controls and relatively complete documentation records. GenScript is capable of providing documents, site audit and other support to help with the applications of your projects in specific regions.

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 %
Purity	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

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

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 scale.

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