CRISPR library screening to develop HEK293-derived cell lines with improved lentiviral vector titers

Lentiviral (LV) vectors have emerged as powerful tools for treating genetic and acquired human diseases. As clinical studies commercial demands progressed, there has been a growing need large amounts of purified LV vectors. To help meet this demand, we developed CRISPR library screening methods to identify perturbations in embryonic kidney 293 (HEK293) cells their derivatives that may increase vector titers. Briefly, vector-based Human Activation Knockout libraries (Calabrese Brunello) were used modify HEK293 HEK293T cells. These cell populations then expanded, integrated genomes rescued by transfection. harvested, the process sequential transduction rescue-transfection was iterated. Through workflow, guide RNAs (gRNAs) target genes suppress or enhance production enriched identified with Next-Generation Sequencing (NGS). Though more work is needed test screen, expect here, such TTLL12 , which an inhibitor antiviral innate immunity be introduced multiplexed yield lines improved productivity.