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...
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...
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 ...
Using a genome-wide CRISPR knock-out screen, the authors defined the repertoire of genes that are required for proerythroblast survival and for terminal erythroid differentiation. Greggory Myers Ann Friedman Rami Khoriaty ResearchOpen Access12 Apr 2025Nature Communications ...
About 15 million reads were obtained for each indexed screen. For some of the screens, an alternative strategy was used to accommodate sequencing on an Illumina NextSeq 500 platform. Changes to the protocol above include amplification of extracted gDNA by PCR using a barcoded forward primer (CC_...
ResearchOpen Access18 Apr 2025Nature Communications Volume: 16, P: 3709 A genome-wide screen identifies genes required for erythroid differentiation Using a genome-wide CRISPR knock-out screen, the authors defined the repertoire of genes that are required for proerythroblast survival and for terminal...
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...
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...