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基因组编辑工具 瑞典皇家科学院决定将2020年诺贝尔化学奖授予埃马纽埃尔·卡彭蒂耶（Emmanuelle Charpentier）和詹妮弗·杜德纳（Jennifer A Doudna），以表彰她们为基因组编辑方法开发所做出的贡献（for the development of a method for genome editing）。主要应对stress conditions，例如heat、nutrient starvation，他们可被压力调节器RpoE2控制。 NHEJ不仅可以修复DSBs，还可能会在breaks处添加外源DNA，也就是说可以促进水平基因转移。 需要注意的是，重组是指来源不同的DNA组合，但重组不一定导致水平基因转移。这篇文章介绍了， 利用Cas9在基因组定点位置打开一个缺口，再通过HITI（非同源末端连接 (NHEJ)的增强版）将目的基因插入。而HITI的优势在于，既可以在分裂的细胞中发生，也可以在非分裂的细胞中发生，同时效率也非常高，这也就是这篇文章的意义所在—— 疾病的治疗，高效且不被细胞类型限制 --- zhihu com question 665881922biology stackexchange com does-non-homologous-end-joining-always-result-i biology stackexchange com questions 38828 how-does-nhej-cause-indels通过创造性地连接tracrRNA-crRNA为嵌合RNA——sgRNA（single guide RNA），简化的操作过程：sgRNA的在PAM的参与、Cas9的介导下与双链DNA配对，Cas9的2个活性中心切割靶标DNA为DSB，引起NHEJ机制，产生突变。 sgRNA的tracrRNA段越长，Cas9的切割效果越好。End Resection Independence: Unlike HR, NHEJ does not require extensive end resection of DNA ends before repair can occur This allows NHEJ to function effectively even when the cell is not actively dividing or when DNA ends are damaged, which is often the case following exposure to ionizing radiation or oxidative stress --- zhihu com question 459350551我需要手动设计，因此一些专门设计gRNA的网站对我用处不大，我就是想系统的学习一下这个过程。I was reading about non-homologous end joining (NHEJ) in my molecular biology of the gene textbook but the explanation provided in the text was rather vague to me, and I was not able to understand itNHEJ 的过程大致如下： 末端识别和结合： Ku70 Ku80 异二聚体识别并结合到 DNA 双链断裂 (DSB) 的两端。 复合体形成： Ku 蛋白招募 DNA-PKcs 激酶和其他辅助因子（如 Artemis 核酸酶、聚合酶 μ 和 λ ），形成一个稳定的复合体，将 DNA 断裂的两端固定在一起。--- zhihu com question 265204609--- zhihu com question 390784251biology stackexchange com how-does-non-homologous-end-joining-nhej-workI was reading up on CRISPR-cas9 and how it works and I am having trouble wrapping my head around how NHEJ to repair the DSB can cause indels to occur Shouldn't the NHEJ just stick the two strands --- zhihu com tardis zm art 656487871NHEJ过程中所需的蛋白Ku70-Ku80以及Rad1可以抑制MMEJ路径，当这些蛋白不再与DNA损伤部位结合时，MMEJ路径被激活，此时MRX复合物、Sae2以及Exo1通过核酸酶作用从5'至3'端切除损伤部分的序列，然后暴露出微同源序列。--- zhihu com question 52820736--- zhihu com question 410759583
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