For those who want to screen such cell lines, and particularly neurospheres, we provide a step-by-step description of assay development tests to be performed before screening, as well as for the screen itself. We highlight considerations of variables that make these screens distinct from, or ...
然而,单纯的设计一条或多条sgRNA(single guide RNA)进行基因敲低,这样的实验方法通量过低,因此,借鉴shRNA Screen技术,许多团队开发出版本各异的Cas9 Screen技术。 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,...
protocol for genome-scale knockout and transcriptional activation screening using the CRISPR-Cas9 system. Custom- or ready-made guide RNA libraries are constructed and packaged into lentiviral vectors for delivery into cells for screening. As each screen is unique, we provide guidelines for determining...
et al. A direct in vivo RNAi screen identifies MKK4 as a key regulator of liver regeneration. Cell 153, 389–401 (2013). Article CAS PubMed Google Scholar Rogers, Z. N. et al. A quantitative and multiplexed approach to uncover the fitness landscape of tumor suppression in vivo. Nat....
By using MIC-Drop, a typical screen of 100 genes can be conducted within 2-3 weeks by a single user. This protocol for Multiplexed Intermixed CRISPR Droplets uses microfluidics to create droplets containing Cas9, multiplexed single-guide RNAs and corresponding DNA barcodes, allowing large-scale ...
CRISPR interference (CRISPRi) is a powerful tool to link essential and nonessential genes to specific phenotypes and to explore their functions. Here we describe a protocol for CRISPRi screenings to assess genome-wide gene fitness in a single sequencing
A multiparametric anti-aging CRISPR screen uncovers a role for BAF in protein synthesis regulation NGPS is an accelerated aging condition, caused by a mutation in BAF. Here, we report the results of an unbiased, whole genome CRISPR microscopy screen aimed at restoring nuclear envelope integrity in...
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 an effic...
(clustered regularly interspersed short palindromic repeat-CRISPR associated nuclease 9) knockout (KO) screen is a powerful and unbiased approach to identify genes that, when deleted, confer PARPi sensitivity or resistance. Previous studies using CRISPR screens in non-PCa cell lines have identified ...
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Extended data figures and tables Extended Data Fig. 1 Mevalonate pathway effects, co-culture screen consistency, and gene set enrichment analysis (GSEA). (a) Schematic...