This chapter discusses how the CRISPR sequences were discovered in bacterial and archaeal genomes, the basis of their classification, the molecular mechanism that helps in genome modification, the applications of CRISPR-Cas9-mediated genome editing, and the challenges that this technology needs to ...
CRISPR is becoming an indispensable tool in biological research. Once known as the bacterial immune system against invading viruses, the programmable capacity of the Cas9 enzyme is now revolutionizing diverse fields of medical research, biotechnology, an
在之后的研究中，研究人员通过解析Cas9的物理化学结构、Cas9切割双链的作用机制等继续赋予cas9新的功能。比如通过突变Cas9的结构域得到了nCas9、dCas9，将他们与一些效应因子（如转录调节因子、DNA甲基化修饰因子、脱氨酶、逆转录酶等）融合开发了新的调控工具：转录激活/抑制系统、单碱基编辑器（如胞嘧啶碱基编辑器-C...
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ii. CRISPR-Cas9 Expression The levels as well as the duration of Cas9 expression are highly critical for CRISPR-Cas9 based genome editing. In stable clones, high expression levels of Cas9 may result in non-specific activity. This may be controlled through isolating multiple clones and screening ...
RNA-guided human genome engineering via Cas9. Science 339, 823 (2013). CAS PubMed PubMed Central Google Scholar Wang, H. et al. CRISPR/Cas9 in genome editing and beyond. Annu. Rev. Biochem. 85, 227–264 (2016). CAS PubMed Google Scholar Sternberg, S.H. & Redding, S. DNA ...
Doudna 因发现 CRISPR/Cas9 基因剪刀,而获得 2020 年诺贝尔化学奖。 CRISPR/Cas 系统由一小段 RNA 和一种高效的 DNA 切割酶 (Cas 核酸酶) 组成的系统,该技术不像 TALENs 技术和 ZFNs 技术是依赖于蛋白与靶基因之间的识别,而是由 sgRNA 和靶基因之间形成复合物,从而完成特定基因序列的编辑。 CRISPR/Cas9 技术...
Morita S, Noguchi H, Horii T, et al. Targeted DNA demethylation in vivo using dCas9-peptide repeat and scFv-TET1 catalytic domain fusions.Nat Biotechnol.2016, 34(10):1060–5. Moradpour M, Abdulah SNA. CRISPR/dCas9 platforms in plants: strategies and applications beyond genome editing.Plant...
Editing endogenous genomic DNA in eukaryotic cells is now possible with CRISPR-Cas9 technology. By taking advantage of a cell’s own DNA repair mechanisms, we are able to add, change, or delete almos...
Morita S, Noguchi H, Horii T, et al. Targeted DNA demethylation in vivo using dCas9-peptide repeat and scFv-TET1 catalytic domain fusions. Nat Biotechnol. 2016, 34(10):1060–5. Moradpour M, Abdulah SNA. CRISPR/dCas9 platforms in plants: strategies and applications beyond genome editing. ...