recombinant
expressed in E. coli
Quality Level
assay
≥95% (SDS-PAGE)
form
lyophilized powder
packaging
pkg of 1 kit (3 components)
application(s)
CRISPR
shipped in
wet ice
storage temp.
−20°C
General description
来源于 化脓性链球菌 的重组Cas9蛋白 (~160KD)是一种用于基因组工程实验的即用型试剂。当与靶标特异性的向导RNA结合时,野生型 化脓性链球菌 Cas9蛋白可作为靶向核酸酶,用于细胞培养物的转染和通过单细胞胚胎注射加速遗传修饰动物的发育。
CAS9,也称为cas5和csn1,是II型聚类规则间隔短回文重复序列(CRISPR)-RuvC(RNase H样折叠)cas系统的特征基因。CAS9含有1388个氨基酸。经预测,该蛋白质含有可参与crRNA成熟的RuvC/核糖核酸酶(RNase)H结构域和可参与DNA降解步骤的McrA/HNH特征结构域。
Application
功能基因组学/靶标验证/基因组编辑
Biochem/physiol Actions
CRISPR/Cas被细菌和古细菌用作防御入侵病毒和质粒的防御系统。最近,来源于 化脓性链球菌 的II型CRISPR/Cas系统已被设计为在真核系统中起作用,其使用两种分子组分:单一Cas9蛋白和非编码向导RNA(gRNA)。可以通过gRNA对Cas9内切核酸酶进行编程,从而将DNA双链断裂(DSB)导向所需的基因组位置。与由锌指核酸酶(ZFN)诱导的DSB类似,细胞随后激活内源DNA修复过程[非同源末端连接(NHEJ)或同源定向修复(HDR)],以愈合目标DSB。
CAS9在质粒DNA干扰中起着至关重要的作用。它是唯一对外源DNA具有抗性的cas蛋白。CAS9可刺激RNA向导的基因组编辑和各种生物体中的基因调控,但它可以在任何给定细胞内一次仅促进一种活性。
Features and Benefits
- 高度特异
- 高活性
- 可直接进行注射/转染
Packaging
50μg包装(≥300pmol)
250μg包装(≥1500pmol)
250μg包装(≥1500pmol)
Preparation Note
冻干化脓性链球菌Cas9蛋白应重悬于所提供的重构溶液,获得所需浓度。轻轻敲打试管,完全溶解冻干粉末,在冰上孵育10分钟,然后离心试管使内容物沉淀至试管底部。
Other Notes
每个试剂盒包括:
- 一瓶Cas9重组蛋白
- 一瓶1×稀释缓冲液(1mL)
- 一瓶含甘油的无核酸酶水(1mL)
仅试剂盒组分
产品编号
说明
- Cas9-NLS from Streptococcus pyogenes, expressed in Escherichia coli
- Dilution buffer for Cas9 proteins
- Reconstitution solution for Cas9 proteins
存储类别
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
wgk
WGK 3
实验方案
可靠的PURedit™ CRISPR合成gRNA和Cas9蛋白可提供行业领先的靶位点切割和特异性
Guaranteed PURedit™ CRISPR synthetic gRNAs and Cas9 protein offer industry-leading on-site cutting and specificity
Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.
商品
Validate CRISPR gene editing experiments easily with Sigma-Aldrich® T7E1 mismatch detection kit, ensuring successful editing.
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