CRISPR Screening in Primary T lymphocytes - A Masterclass
Expanding CRISPR screening technology to ex vivo primary T cells through a combined lentiviral and electroporation approach
8 Feb 2019
Expert insights
Genetic screens are powerful tools to interrogate cell responses to drug treatment as well as microenvironment perturbations. With the increasing interest in manipulating the immune system to tackle disease and the use of CRISPR (clustered regularly interspaced short palindromic repeats)–Cas9 genome editing systems, screening primary cells is both appealing and feasible
In an upcoming webinar, Dr. 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.
Ghirelli will cover key points of CRISPR screening in primary human T cells, including:
- Human T cell gene editing
- Donor variability in T cells CRISPR screening
- NGS data analysis for genetic screenings
Who should attend:
Scientists with a specific interest in CRISPR screening in primary human cells, looking for CRISPR technology optimization or those with a particular interest in T cell target identification.
Attend the live webinar:
The live webinar will take place on Thursday, February 28, 2019 at:
- 15:00 GMT
- 10:00 EST
- 07:00 PST
- 16:00 CET
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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