Figure 2.Workflow for lentiviral pooled screening.A Cas9-expressing stable cell line is generated with LentiArray Lentiviral Cas9 Nuclease using selection for blasticidin resistance. These Cas9-expressing cells are then transduced with the LentiPool sgRNA library at the appropriate MOI and undergo select...
Transduction with pooled sgRNA library Lentiviral transduction of Cas9-expressing cells with LentiPool sgRNA library Selection with puromycin Expansion of resistant cells Primary screening by positive or negative selection: Positive selection (+): Apply treatment to cells (e.g....
目录 CRIPSR/Cas9 Screening by GeCKO-V2 1. 评估GeCKO-V2质粒文库的好坏 样本比对信息统计表 gRNA的read count 分布图 均一性斜率图 2 利用MageCK分析CRISPR文库筛选数据 下载CRIPSR-Screening library list 合并双端测序fastq文件 计算样本中gRNA的均一化counts 使用Mageck mle计算不同样本间组合情况 评估GeCKO-V2...
CRISPR/Cas9-based tools have rapidly developed in recent years. These include CRISPR-based gene activation (CRISPRa) or inhibition (CRISPRi), for which there are libraries. CRISPR libraries for loss of function have been widely used to identify new biological mechanisms, such as drug resistance ...
1.Kurata Morito,et al. CRISPR/Cas9 library screening for drug target discovery.[J] .J. Hum. Genet., 2017. 2.Wang Tim,et al. Genetic screens in human cells using the CRISPR-Cas9 system.[J] .Science, 2014, 343(6166): 80-4.
讲了这一些,似乎本人很不推荐稳转这种方法?其实这依据个人的实验目的选择。那些需要有稳定基因背景的研究来说,最好的选择是快敲不残留的方法(瞬转)。对于追求速度,不拘泥于什么基因敲除效果,尤其是文库筛选(CRISPR-Cas9 library screening),则使用慢病毒的方式是相对适宜的。选用哪种方式,需要多多考量。
Sorafenib is the standard treatment for advanced hepatocellular carcinoma (HCC). However, the development of drug resistance is common. By using genome-wide CRISPR/Cas9 library screening, we identify phosphoglycerate dehydrogenase (PHGDH), the first committed enzyme in the serine synthesis pathway (SSP...
[4] Deng L, Yang L, Zhu S, et al. Identifying CDC7 as a synergistic target of chemotherapy in resistant small-cell lung cancer via CRISPR/Cas9 screening[J]. Cell Death Discovery, 2023, 9(1): 40. [5] Tang S, Wu X, Liu J, et al. Generation of dual-gRNA library for combinatoria...
全基因组CRISPR/Cas9高通量筛选人体细胞中的功能基因组学 英文名:《Genome-Wide CRISPR/Cas9 Screening for High-Throughput Functional Genomics in Human Cells》 简单说就是利用CRISPR/Cas9技术对人类的基因组(23对染色体上的全部基因,大约两万个)...
Transduction with pooled sgRNA library Lentiviral transduction of Cas9-expressing cells with LentiPool sgRNA library Selection with puromycin Expansion of resistant cells Primary screening by positive or negative selection: Positive selection (+): Apply treatment to cells (e.g., drugs, chemical p...