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Cas9 Geminin plasmid

品牌
Sigma-Aldrich
货号
CAS9GEMP
包装型号
规格纯度
参考价格
2486 *本价格含增值税费
促销
服务
  • 原厂保证
  • 包邮
  • 增值税票
数量
-+
产品名称:
Cas9 Geminin plasmid
产品介绍:

产品说明

一般描述

This product is an expression plasmid that utilizes the EF1a promoter for strong transient expression of a Cas9-GFP-Geminin fusion (EF1a-Cas9-GFP-Geminin) allowing for easy visualization of successful transfection. The Cas9-Geminin expression plasmid is one part of a two part CRISPR system with individual Cas9-Geminin and gRNA expression vectors.

To order gRNA in any format click here

应用

Functional Genomics/Target Validation

  • Performing HDR mediated targeted integration in multiple cell lines
  • Creation of knock-in cell lines with promoters, fusion tags or reporters integrated into endogenous genes

特点和优势

  • Highly specific and highly active
  • Sequence verified
  • 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. While HDR is absent in G1, NHEJ is active throughout the cell cycle and is largely favored over HDR. Consequently HDR can be increased by directly synchronizing the expression of Cas9 with cell-cycle progression by fusing Cas9 to human Geminin (a protein expressed in S and G2 phases). The Cas9-geminin fusion protein then regulates gene editing by promoting repair during S and G2 phases when homology directed repair (HDR) occurs. The result is creation of double strand breaks in cells at times that are more able to incorporate donor sequences.

法律信息

CRISPR Label License

产品性质

重组expressed in E. coli
包装vial of 50 μL
浓度20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
启动子Promoter name: EF1-alpha
application(s)CRISPR
报告基因GFP
selectionampicillin
运输dry ice
储存温度−20℃

安全信息

储存分类代码12 - Non Combustible Liquids
WGKWGK 2
闪点(F)Not applicable
闪点(C)Not applicable

Sigma-Aldrich

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