GeneHero™ CRISPR human sgRNA libraries

Introduction

GeneCopoeia offers CRISPR sgRNA libraries for high-throughput knockout of human genes in specific or custom gene groups or pathways. Loss-of-function screening by gene knockout is a powerful tool for systematic genetic analysis in mammalian cells, facilitating gene discovery, genome-scale functional interrogation (e.g., signal transduction pathways) and drug discovery (e.g., target identification and drug mechanism studies).

The GeneHero™ human single guide RNA (sgRNA) libraries are individually cloned into lentiviral vectors and sequence-verified for dual-use (transfection or transduction) delivery methods designed for large-scale functional screens. For each targeted gene, a minimum of 2 barcoded sgRNAs targeting different regions are created, optimized and sequence-verified to ensure efficient gene knockout. Further, each sgRNA-expressing plasmid is individually cultured in E. coli before pooling, providing the best possible representation of each sgRNA in the pools. The libraries can be ordered as pools of sgRNAs in pre-defined gene families or as custom sets, and are available as lentiviral particles, transfection-ready plasmid DNA or bacterial stocks. They can be transfected or transduced into cell lines, including those stably expressing Cas9 nuclease.

Illustration-of-CRISPR_Cas9-mediated-genome-editing

 

Figure 1. Illustration of large-scale screening with sgRNA library

 

Applications

  • High-throughput knockout screening with many sgRNAs, either individually or in pools.
  • Drug target discovery (Figure 2).
  • Drug target validation.
  • Phenotypic screens.
  • Reporter assays.

 

 

Figure 2. Workflow for CRISPR sgRNA libraries. A. Pooled screen. Cells infected with each sgRNA library pool are screened for the desired readout. Pooled cells are subjected to Sanger sequencing for individual sgRNAs, or deep sequencing to look for over- or under-representation of individual sgRNAs. B. Knockout screen using arrayed sgRNAs. Cells are infected with individual sgRNA lentiviruses. Wells are screened for the readout of interest. Individual sgRNAs corresponding to the phenotype of interest are already known without sequencing.