The Pooled Lentiviral Human Genome-Wide CRISPR gRNA Library is the latest tool for performing loss-of-function screens. The CRISPR/Cas9 system can be applied for permanent gene knockout by using a combination of gRNA and a Cas9 nuclease.
Target cells are transduced with Cas9-expressing lentivirus and selected to optimal Cas9 expression levels. The Cas9 activity level can be assessed using the CRISPRtest Functional Cas9 Activity Kit. The pooled gRNA library is then transduced into cells so that a majority of the cells receives only one gRNA construct. Loss-of-function screening followed by next generation sequencing (NGS) of integrated gRNA from genomic DNA reveals which genes are involved in the biological process studied.