The promoter used in this product is CVM which is designed for use with Mammalian systems. This product has not been tested outside of Mammalian systems.
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重组
expressed in E. coli
质量水平
包装
vial of 50 μL
浓度
20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
运输
dry ice
储存温度
−20°C
一般描述
This product is an expression plasmid that utilizes the CMV promoter for strong transient expression of eSpCas9 and GFP linked by a 2A peptide (CMV-eSpCas9-2A-GFP) allowing for easy visualization of successful transfection. The eSpCas9 expression plasmid is one part of a two part CRISPR system with individual eSpCas9 and gRNA expression vectors.
To order gRNA in any format click here
To order gRNA in any format click here
应用
Functional Genomics/Target Validation
- Creation of gene knockouts in multiple cell lines
- Complete knockout of genes not amenable to RNAi
- Creation of knock-in cell lines with promoters, fusion tags, or reporters integrated into endogenous genes
特点和优势
- Enhanced specificity compared to wild type Cas9
- Highly Active
- Ready to use purified plasmid DNA
原理
CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB. Newly engineered eSpCas9 enables the efficient targeted gene editing of established CRISPR systems with the benefit of reduced off-target effects. Point mutations in the chromosome-binding motif of SpCas9, as described by Slaymaker, et al., provide higher on-target fidelity without loss of cleavage efficiency.[1]
储存分类代码
10 - Combustible liquids
WGK
WGK 2
闪点(°F)
Not applicable
闪点(°C)
Not applicable
其他客户在看
Rationally engineered Cas9 nucleases with improved specificity.
Slaymaker, I.M., et al.
Science, 351, 84-88 (2015)
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Is this plasmid suitable for bacteria amplification and metazoan GFP expression?
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