二、CRISPR Screen 2.1protocal 高通量是新时代生物技术的标配,从能一次完成超过5万个基因的PCR(RNA-seq)到细胞谱系追踪(barcoding),在不影响研究精准度的情况下,大大的降低科学研究时间和成本。依据我们前面的介绍,sgRNA将Cas9蛋白靶向至目的序列,Cas9结合到DNA上后剪切DNA,那么以下这几种情况则非常好理解: (1)Cas9...
第三期多组学系列研讨会有幸邀请到西湖大学马丽佳研究员与大家分享CRISPR遗传筛选中的细胞分子表型鉴定技术与应用(Enable high content molecular phenotyping in CRISPR screen),主要会围绕以下几个方面进行探讨。• 使用多重CRISPR筛选鉴定高维基因相互作用(Use mul
Discussion PCa genetic risk SNPs are enriched in noncoding CREs rather than in protein-coding regions9,80. It is challenging to systematically evaluate the importance of these rCREs in cancer biology and the clinic. Our study demonstrates that CRISPRi mediated loss-of-function screen of rCREs is...
CRISPR screens in the field of virology are ideally suited for analyzing virus-host interactions; determining host factors critical for virus entry, replication, and spread; and identifying and prioritizing novel drug targets. Performing a genome-scale CRISPR/Cas9 knockout screen with an sgRNA library...
The cellular processes that govern tumor resistance to immunotherapy remain poorly understood. To gain insight into these processes, here we perform a genome-scale CRISPR activation screen for genes that enable human melanoma cells to evade cytotoxic T c
We demonstrate Orthrus' features for screen quality assessment and two distinct scoring modes for dual guide RNAs (dgRNAs) that target the same gene twice or dgRNAs that target two different genes. Running Orthrus requires basic R programming experience, ~5鈥 10 min of computational time and 15...
十、CRISPR转录抑制/转录激活以及CRISPRscreen 1、dCas9的特征简介 2、运用CRISPR技术进行转录调控的原理 3、CRISPR转录抑制/转录激活技术及优化 4、CRISPRoff 5、CRISPR screen及CRISPRa/iscreen(高通量筛选) 十一、单碱基编辑与单碱基编辑器(CBE、ABE、PE) ...
Somatic loss-of-function CDK12 mutations generate an aggressive subtype of CRPC with poor outcomes [19,20,21]. While we identified CDK12 as critically required for PCa cell survival via a CRISPR screen. Both genetic deletion and functional inhibition suggested that CDK12 accelerates PCa progression...
The gRNA library-expressing cells can be transplanted into an animal model to perform indirect in vivo screens or can be directly used as an in vitro screen model (Figure 2). Additionally, the naked gRNA library can be injected into an animal model to perform direct in vivo screens (Figure...
我查询到的最早版本的Cas9 screen来自两篇背靠背Science paper,分别出自Broad研究所张锋团队和Langer与Sabitini团队[1][2]。在张锋团队发表的这篇文章中,Cas9 screen技术被称为GeCKO(Genome-scale CRISPR-Cas9 Knockout)。首先,该团队利用一个载体表达U6-sgRNA和EFS-SpCas9,减少基因表达不均一性(图1A)。随后,团队比...