Crispr/Cas9 腺病毒

高效极速基因敲除神器-Crispr/Cas9 腺病毒

 

 

      Crispr/Cas9 简介

CRISPR (clustered, regularly interspaced, short palindromic repeats)是一种来自细菌降解入侵的病毒 DNA 或其他外源 DNA 的免疫机制。目前,来自Streptococcus pyogenes 的CRISPR-Cas9系统应用最为广泛。Cas9 蛋白(含有两个核酸酶结构域,可以分别切割DNA 两条单链。Cas9首先与crRNA及tracrRNA结合成复合物,然后通过PAM序列结合并侵入DNA,形成RNA-DNA复合结构,进而对目的DNA双链进行切割,使DNA双链断裂。

CRISPR-Cas9系统已经成功应用于植物、细菌、酵母、鱼类及哺乳动物细胞,是目前最高效的基因组编辑系统。

通过基因工程手段对crRNA和tracrRNA进行改造,将其连接在一起得到sgRNA(single guide RNA)。融合的RNA具有与野生型RNA类似的活力,但因为结构得到了简化更方便研究者使用。通过将表达sgRNA的原件与表达Cas9的原件相连接,得到可以同时表达两者的质粒,将其转染细胞,便能够对目的基因进行操作。

汉恒生物创新性的首次将cas9和gRNA整合到腺病毒载体中,成功包装cas9-gRNA腺病毒,很大程度上弥补了crispr/cas9质粒感染效率低,从而导致敲除效率低的缺陷。

汉恒生物推出的Cas9/gRNA腺病毒利用cas9和gRNA结合特异性,同时借助腺病毒高感染效率,高表达效率,提高crispr/cas9基因敲除效率,缩短基因敲除周期,使得基因敲除周期短,成本低,更易于操作。

Crispr/Cas9腺病毒特点:
   

1、可感染多种分裂期和非分裂期细胞,感染效率高、敲除效率高;

2、操作简单,容易掌握;

3、基因敲除周期短,成本低;

4、适合哺乳动物不同种属不同基因的操作,适用范围广。

 

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