Pooled CRISPR KO Libraries

Pooled Lentiviral CRISPR KO sgRNA Libraries for Genome Wide CRISPR KO Screenings

Cellecta´s genome wide pooled lentiviral CRISPR KO sgRNA libraries contain 69k – 89k defined single-guide RNA sequences enabling unbiased disruption (“knockout”) of all protein coding genes in the genome throughout a large cell population in a single experiment. Screening this cell population for a specific phenotype (e.g., viability) enables the identification of specific genes driving this phenotype of interest, such as a drug response or disease phenotype.

Pooled CRISPR sgRNA Lentiviral Libraries are transduced into Cas9-expressing cells to disrupt thousands of genes simultaneously across different cells in a large population. Selection of cells with a phenotype of interest (e.g., viability or activation of a reporter) then leads to enrichment or depletion of particular sgRNA that target genes functionally important in regulating the phenotype.

On completion of the screen, genomic DNA from the whole population is isolated and the frequency of each integrated sgRNA lentiviral construct in the population is assessed by next-generation sequencing (NGS).

Nucleus Biotech provides Cellecta´s human, mouse, chicken and pig whole genome pooled CRISPR KO libraries as well as separate modules of the human library covering different pathways and approved drug targets. All libraries are offered as plasmid DNA in large quantities enabling lentiviral packaging of the original library without the need for prior plasmid amplification, or as pre-packaged lentiviral particles ready for transduction.

Cellecta Pooled CRISPR KO Libraries – The Advantages:

• HEAT sgRNA scaffold for better Cas9 binding improves screening results
• sgRNAs are designed for maximum efficiency and specificity (Doench et al., 2016)
• Pooled sgRNA sequences are synthesized on Agilent chips ensuring uniform distribution, and subsequently cloned in Cellecta´s highly efficient lentiviral vector backbone
• sgRNA distribution in the library pool is proven by stringent NGS QC: All libraries are delivered with the NGS QC data details demonstrating the distribution of the sgRNA sequences in the pool
• Delivery of 200 ug plasmid DNA pool enabling lentiviral packaging of the original library without the need for prior plasmid amplification, or as pre-packaged lentiviral particles (2 x 10e8 TU or 1 x 10e9 TU) ready for transduction.
• Special pricing for academic institutions