CRISPR-Cas9基因编辑技术简介 - 知乎 - 知乎专栏
CRISPR-Cas9基因编辑技术就是通过人工设计的 sgRNA（guide RNA）来识别目的基因组序列，并引导 Cas9 蛋白酶进行有效切割 DNA 双链，形成双链断裂，损伤后修复会造成基因敲除或敲入等，最终达到对基因组DNA 进行修饰的目的。
只需5步，快速学会CRISPR/Cas9的sgRNA设计 - 知乎 - 知乎专栏
而通过人工设计crRNA和tracrRNA这两种 RNA，可以改造形成具有引导作用的sgRNA （single guide RNA ），从而引导 Cas9 对 DNA 的定点切割。 使用CRISPR/Cas9编辑基因成败的关键就在于sgRNA，在线设计sgRNA的网站有很多，本文重点推荐并介绍一个功能丰富，操作非常简单的站 …
Selection of highly efficient sgRNAs for CRISPR/Cas9-based
2016年2月19日 · Here, we developed two modules to construct CRISPR/Cas9-sgRNA system containing multiple sgRNA cassettes. Both modules employ common restriction enzymes to perform molecular cloning.
CRISPR/Cas9实验实操专题一：sgRNA设计 - 知乎 - 知乎专栏
1.sgRNA设计. 1.1设计原则. 1） II型CRISPR系统（SpCas 9）的sgRNA 靶点一般为20nt. 2）sgRNA序列的碱基组成：基因特异的sgRNA 模板序列位于PAM 序列前，PAM序列的特征为NGG (N可以为任意核苷酸) 3）sgRNA的序列应避免以4个以上的T结尾，GC%含量为30%-70%
Multiplexed CRISPR technologies for gene editing and ... - Nature
2020年3月9日 · Multiplexed CRISPR technologies, in which numerous gRNAs or Cas enzymes are expressed at once, have facilitated powerful biological engineering applications, vastly enhancing the scope and ...
A versatile, high-efficiency platform for CRISPR-based gene
2023年2月17日 · CRISPR-mediated transcriptional activation (CRISPRa) is a powerful technology for inducing gene expression from endogenous loci with exciting applications in high throughput...
DeepCRISPR : optimized CRISPR guide RNA design by deep learning
2018年6月26日 · Here we present DeepCRISPR, an efficient and extendable computational model for simultaneous prediction of CRISPR sgRNA on-target knockout efficacy and whole genome off-target profiles. DeepCRISPR surpassed the state-of-the-art tools across a variety of human datasets with solid evaluation metrics.
SgRNA engineering for improved genome editing and
2022年6月1日 · This review highlights representative studies that optimize the sgRNA nucleotide sequences for improved genome editing performance (e.g. activity and specificity) as well as add extra aptamers and end extensions for expanded CRISPR-based functional assays (e.g. transcriptional regulation, genome imaging, and prime editor).
CRISPR Guide - Addgene
Engineered CRISPR systems contain two components: a guide RNA (gRNA or sgRNA) and a CRISPR-associated endonuclease (Cas enzyme). The gRNA is a short synthetic RNA composed of a scaffold sequence necessary for Cas-binding and a user-defined ∼20-nucleotide spacer that defines the genomic target to be modified.
Optimizing sgRNA structure to improve CRISPR-Cas9 knockout …
2015年12月15日 · By a systematic investigation of sgRNA structure we find that extending the duplex by approximately 5 bp combined with mutating the continuous sequence of thymines at position 4 to cytosine or guanine significantly increases gene knockout efficiency in CRISPR-Cas9-based genome editing experiments.
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