小白快速上手！CRISPR-KO系统的构建 - 知乎 - 知乎专栏
Apr 11, 2023 · CRISPR 最初用于敲除（knock out，KO）各种细胞类型和生物体中的靶基因，但对各种 Cas 酶的修饰使 CRISPR 扩展到选择性激活/抑制靶基因、纯化 DNA 的特定区域、活细胞中的 DNA 成像以及精确编辑 DNA 和 核糖核酸。
A simple method using CRISPR-Cas9 to knock-out genes in murine ... - Nature
Dec 18, 2020 · CRISPR-Cas9 system can be used to generate knock-out cancer cell lines. An insertion or deletion induced by a single guide RNA (gRNA) is often used to generate knock-out cells, however, some...
如何进行CRISPR/Cas9介导的基因敲除的验证？ - 知乎专栏
近期关于CRISPR/Cas9系统做编码基因的KO（敲除）的验证方法问题，收到很多咨询。. 大体分成两个，分别是： 是否可以通过用qPCR检测目的基因的mRNA来验证是否有敲除效果？. 以及是否可以直接用wb验证目的蛋白的表达量来验证是否有敲除效果？. 因此决定写一篇 ...
An improved strategy for CRISPR/Cas9 gene knockout and …
Feb 13, 2020 · The gene knockout (KO) efficiency of CRISPR/Cas9 mainly depends on cellular Non-homologous End Joint (NHEJ) repair to maintain genomic integrity if a complementary donor is not present when Cas9 cleaves both double strands at gRNA binding sites.
Genome-scale CRISPR-Cas9 knockout and transcriptional
Dec 10, 2014 · As a proof of principle to demonstrate the CRISPR-Cas9 system's utility for screening, we constructed genome-scale CRISPR-Cas9 knockout (GeCKO) and SAM libraries to identify genes that,...
如何利用CRISPR/Cas9构建稳定敲除细胞系？ - 安必奇生物科技
如何利用CRISPR/Cas9构建稳定敲除细胞系？. 今天我们聊聊如何利用CRISPR/Cas9技术进行 稳定基因敲除细胞系的构建，以293T细胞为例。. sgRNA验证：在实验开始前，可以通过sgRNA在线验证，筛选高效率的sgRNA用于敲除细胞系的构建。. 基因拷贝数：建议在开始实验之前 ...
An efficient and precise method for generating knockout cell lines ...
Sep 7, 2020 · Here, we adapted a targeted knock‐in strategy with CRISPR‐Cas9 system and characterized the efficiency for generating single or double knockout cell lines. Specifically, a homology‐arm based donor cassette consisting of genes encoding a fluorescence protein and antibiotic selection marker driven by a constitutive promoter was co ...
CRISPR–Cas9 screens reveal regulators of ageing in neural
Oct 2, 2024 · To systematically identify genes that boost NSC activation as a function of age, we developed a genetic screening platform to conduct genome-wide CRISPR–Cas9 knockout screens in primary NSC...
Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing
Aug 21, 2021 · CRISPR and its associated protein (Cas-9) is a method of adaptive immunity in prokaryotes to defend themselves against viruses or bacteriophages. 2 Japanese scientist Ishino and his team accidentally found unusual repetitive palindromic DNA sequences interrupted by spacers in Escherichia coli while analyzing a gene for alkaline phosphatase first...
CRISPR-Cas9应用——基因敲除与敲入系统 - 知乎 - 知乎专栏
crispr-cas9基因敲入系统是cas9内切酶切割双链dna，且有一段高度同源的dna修复模板存在的情况下，生物体内启动hdr修复路径，将一段外源dna定点插入基因内部。
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