Category: Assay Development and Screening
Target identification leading to novel druggable targets for therapeutic intervention remains a top priority for pharma/biotech industry. CRISPR-based technologies have revolutionized gene function studies and genetic screens to unprecedented levels. Although pooled libraries are an attractive solution for screening large sets of genes at once, they require time-consuming next-generation sequencing (NGS), data analysis, and extensive secondary screens and validation. Discussed here are complete workflow solutions for functional genomics and genome wide screening applications using our award-winning LentiArray™ CRISPR Libraries, and downstream validation assays using TrueGuide™ synthetic guide RNA. LentiArray™ CRISPR libraries are delivered in 96-well format with up to 4 sequence-verified distinct gRNA constructs per gene per well. The CRISPR gRNA sequences were designed using our de novo design algorithm targeting over 18,000 coding genes within human genome. The high gene editing efficiencies of these gRNAs by either transient transfection or viral delivery was demonstrated using NGS and Genomic Cleavage assays. Arrayed screens not only allow to detect cell viability, specific markers and reporters, but also enable one to perform complex phenotypic read outs such as morphology changes, high content imaging, and multiple gene knockouts in a single screen. We show functional validation data for CRISPR libraries using different cell-based assays in arrayed format. These new reverse genetic screening tools serve as powerful platforms for target discovery and validation thereby enabling disease research.
Jian-Ping Yang– Staff Scientist, Thermo Fisher Scientific, Carlsbad, CA
Thermo Fisher Scientific
R&D technical lead for developing tools and products for genome and cell engineering applications within Synthetic Biology group at Thermo Fisher Scientific. Obtained PhD in Molecular Genetics at Nagoya City University Medical School, Japan, and postdoctoral training in University of California San Diego.