CRISPR Knockout Screening

Knockout thousands of genes – all at once.

The discovery of CRISPR genome editing has provided a novel and highly efficient way to probe gene function through the generation of gene knockouts.

Horizon’s CRISPR KO screening service is using a high performance vector system. The all-in one vector system offers significantly improved performance in a rapid workflow (Cross et al., 2016). Our optimized screening platform, sophisticated bioinformatics pipeline and multiple ready-made and custom libraries allow us to design the most appropriate workflow for your screening project to provide you with data of exceptional quality.

CRISPR KO screens are ideally suited to

  • Identify and prioritize drug targets
  • Find genes required for cell viability, drug sensitivity or resistance
  • Guide patient selection for clinical trial
  • Identify targets or pathways for potential combination therapies

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CRISPR KO Screening Platform

Horizon can provide complete workflows for CRISPR knockout screens, from cell line selection and optimisation through to bioinformatics analysis of the screen results.

Turnaround time: 12-20 weeks

Deliverables: Data in raw and analysed form and a final report containing experimental details and hit nominations

Select one of our off-the shelf sgRNA libraries or have us design a custom library!

  • Cell surface, Signalling & Metabolic
  • Deubiquitinases & Drug Targets
  • Kinases & DNA Damage Response
  • Epigenetics & Autophagy
  • GeCKOv2 whole genome library

To learn more about our CRISPR screening platform, watch our recorded webinar:

CRISPR Screening, the What, Why and How

Case study: Whole genome CRISPR KO resistance screen in A375 melanoma cells

Purpose: A proof of concept study to identify resistance factors against a BRAF kinase inhibitor Vemurafenib (PLX-4032) in A375 melanoma cells that carry a BRAF V600E gain-of-function mutation.

Methods:

  • GeCKO v2 knockout library: 6 sgRNAs against 19,050 genes (Sanjana et al., 2014)
  • Pooled-based approach: lentivirus transduction of the library into cells with antibiotic selection
  • Treatment of the edited cell population with Vemurafenib for 14 days
  • Cell pellet collection and sample preparation
  • Next-generation sequencing and data analysis using an adapted MAGeCK workflow

Results:

  • Highest ranking genes were MED12, NF1, CUL3, NF2, TADA2B and TADA1, which are known to  confer resistance to Vemurafenib.1
  • Additional hits include members of the STAGA histone acetyl transferase complex (TAFL5/PAF65b) and the Mediator complex (MED23).

CRISPR Cas9 screen in A375 melanoma cells

Figure 1. Ranking of the hits of the screen by MAGeCK algorithm

Download the complete application note

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