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Merck

CAS9PROT

Sigma-Aldrich

Cas9蛋白

from Streptococcus pyogenes, recombinant, expressed in E. coli, 1X NLS

别名:

Cas9, SpCas9, SpyCas9

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About This Item

分類程式碼代碼:
12352200

重組細胞

expressed in E. coli

品質等級

化驗

≥95% (SDS-PAGE)

形狀

lyophilized powder

包裝

pkg of 1 kit (3 components)

應用

CRISPR

運輸包裝

ambient

儲存溫度

−20°C

一般說明

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。
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。

應用

功能基因组学/靶标验证/基因组编辑

生化/生理作用

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。

特點和優勢

  • 高度特异
  • 高活性
  • 可直接进行注射/转染

包裝

50μg包装(≥300pmol)
250μg包装(≥1500pmol)

成分

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)

原則

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。

重構

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。

法律資訊

MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany

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

儲存類別代碼

10 - Combustible liquids

水污染物質分類(WGK)

WGK 3

閃點(°F)

Not applicable

閃點(°C)

Not applicable


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Yu Shirai et al.
Cell reports methods, 2(5), 100215-100215 (2022-06-01)
Current approaches for insect gene editing require microinjection of materials into early embryos. This severely limits the application of gene editing to a great number of insect species, especially to those whose reproduction systems preclude access to early embryos for
Joseph Andrew Whitley et al.
Journal of extracellular vesicles, 11(4), e12196-e12196 (2022-04-07)
CRISPR/Cas9 genome editing is a very promising avenue for the treatment of a variety of genetic diseases. However, it is still very challenging to encapsulate CRISPR/Cas9 machinery for delivery. Protein N-myristoylation is an irreversible co/post-translational modification that results in the
The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli
Sapranauskas R, et al.
Nucleic Acids Research, 39(21), 9275-9282 (2011)
Orthogonal Cas9 proteins for RNA-guided gene regulation and editing
Esvelt K M, et al.
Nature Methods, 10(11), 1116-1116 (2013)
Ana Carolina Seni-Silva et al.
BMC genomics, 23(1), 123-123 (2022-02-14)
The Natterin protein family was first discovered in the venom of the medically significant fish Thalassophryne nattereri, and over the last decade natterin-like genes have been identified in various organisms, notably performing immune-related functions. Previous findings support natterin-like genes as

商品

After you have performed a CRISPR experiment it is important to determine which gRNAs performed successfully editing. There are many ways to validate CRISPR gene editing experiments. A quick and easy way to check for cutting is by using the Sigma-Aldrich® T7E1 mismatch detection kit.

实验方案

Sigma-Aldrich® sgRNA, crRNA and tracrRNA come HPLC purified as standard with our performance guarantee—even for custom designs.

The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system was discovered in bacteria, where it functions as an adaptive immune system against invading viral and plasmid DNA.

Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.

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