CRISPR Screening in Primary T Cells
1 Mar 2019These slides detail the discussion from the Horizon Discovery webinar on CRISPR (clustered regularly interspaced short palindromic repeats) screening in primary T cells.
In these slides from the webinar, Cristina Ghirelli, immuno-oncology manager at Horizon Discovery, discusses the development of a pipeline to perform pooled CRISPR screening in primary T cells through a combined lentiviral and electroporation approach. With a protocol validated using culture settings previously published in Jurkat T cells (Birsoy et al., 2015), Horizon Discovery treated primary T cells were used to identify new metabolic targets involved in cell proliferation control and survival. Next-generation sequencing data not only confirmed some Jurkat hits, but also revealed new specific T cell targets.
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CRISPR Screening Services
CRISPR Screening Services Horizon offers comprehensive CRISPR screening services from screen design and cell line selection to sophisticated bioinformatics analysis of the screen results. Our best-in-class CRISPR KO (knockout), CRISPRi (interference) and CRISPRa (activation) screening platforms can significantly advance our customers’ drug development programs by offering the highest quality and highest confidence in their screening results. Data analysis services, including sample preparation, Next-Gen Sequencing, and hit calling, can be provided for users of Edit-R Lentiviral sgRNA Pooled Screening Libraries. Simply provide whole cell pellets for each of your samples and we’ll do the rest. We can also help with the NGS data analysis and hit calling for data created using the Edit-R Pooled sgRNA Indexing PCR and Sequencing Primer Kits. Types of CRISPR Screen Expertise includes whole-genome and custom targeted drug resistance and sensitivity screens, synthetic lethality target discovery screens and complex phenotypic screens using a FACS-based readout. We have optimized 50+ cell lines for CRISPR screening and have completed over 300 screens to date. We offer a choice of CRISPR screen types depending on the expression effect you require: CRISPR KO screening with complete loss of gene expression provides the maximal window for phenotypic effect and high statistical power for hit discovery CRISPRi screening represses expression rather than completely knocking out the target gene and is ideally suited to study druggability and to evaluate the function of genes that when knocked-out are essential or that are amplified CRISPRa screening amplifies gene expression in its endogenous context and enables for the first time, to study activation-linked responses on a genome-wide level Dual CRISPRi & CRISPRa screening