Genome-wide knockout screens

Genome-wide CRISPR KO screens with the Inzolia library

The in4mer CRISPR/enCas12a multiplex knockout platform represents a significant advancement in genetic perturbation technology, offering efficient single-gene knockout and genetic interaction screening capabilities. This system utilizes arrays of four independent guide RNAs (gRNAs) targeting either the same or different genes, delivered via an all-in-one lentiviral vector. The genome-scale library (called Inzolia) that targets approximately 19,700 single genes and over 4,900 paralog combinations (pairs, triples, and quads) while being 30% smaller than typical CRISPR/Cas9 libraries. In validation screens across multiple cell lines, the platform demonstrated high sensitivity in identifying core and context-dependent essential genes comparable to CRISPR/Cas9 libraries, while also effectively detecting synthetic lethal and buffering genetic interactions between paralogs. The in4mer paper was published By the Hart and Doench labs in Anvar et al. (Nat. Comm., 2024)