Cas9 screen技术是一项极具潜力的高通量筛选工具,已被应用于各个领域。同时,其衍生与拓展技术,如Cas13 screen、Base editor screen也逐步问世。我相信未来的研究手段一定是从杂乱走向整合、从非系统性走向系统性、从低通量走向高通量的,因此类似Cas9 screen的技术在未来一定大有可为。 参考 ^Shalem, Ophir et al. ...
To identify novel host factors required for HIV-1 infection, we performed a genome-wide CRISPR/Cas9 screen using HIV-induced cell death as a partitioning method. We created a gene knockout library in TZM-GFP reporter cells using GeCKOv2, which targets 19,050 genes,...
2. Add polybrene into DMEM at a final concentration of 8 μg/mL. Infect cells with Cas9-producing virus. 3. Add 5 μg/mL of Blasticidin onto cells 48 h after virus infection to enrich Cas9-expressing cells. Isolate the be...
Thus, CRISPR/Cas9 screen with a hepatitis virus infection background may offer insight into the mechanism for other similar viruses. As a multi-risk factor-induced cancer, the number of CRISPR/Cas9 screens with different backgrounds is still rather limited. The involvement of other HCC risk ...
Antibody–drug conjugates (ADCs) selectively deliver chemotherapeutic agents to target cells and are important cancer therapeutics. However, the mechanisms by which ADCs are internalized and activated remain unclear. Using CRISPR-Cas9 screens, we uncover many known and novel endolysosomal regulators as mod...
Virus supernatant was added to target cells at 48 and 72 hr post-transfection. Target cell selection was then begun 48 hr post-transduction. CRISPR/Cas9 Mutagenesis Studies Following the Avana screen, specific human genes were targeted by CRISPR/Cas9 engineering, using Avana sgRNA sequences. These...
Fig. 2: Enriched irradiation-resistant genes from the genome-scale CRISPR/Cas9 activation screen in GBM. A Significantly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways for genes with sgRNA diversity >3 in irradiation-resistant GBM cells. B The genes with the top 10% of sgRNA...
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)进行筛查,看看这些基因到底有哪些功能。
, 易国强, 唐中林. 基于CRISPR/Cas9 系统在全基因组范围内筛选功能基因及调控元件研究进展. 遗传, 2020,42(5): 435–443. [2] Cai P, Otten ABC, Cheng B, Ishii MA, Zhang W, Huang B, Qu K, Sun BK. A genome-wide long noncoding RNA CRISPRi screen identifies PRANCR as a novel regulator ...
Genome-wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication J. Han, J. T. Perez, C. Chen et al., "Genome-wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication," Cell Reports, vol. 23, no. 2, pp. 596-607, 2018......