While significant progress has been made carrying out CRISPR screens in immortalised cell lines, a more physiological and clinically relevant alternative is human primary cells, such as T cells, regulatory B cells, or natural killer (NK) cells. The use of phenotypically relevant primary cells, though, is not without biological and technical challenges, especially when their intended use includes gene editing and scale-up of these specialised cell types for screening. Verena Brucklacher-Walder at Horizon Discovery analyses the significant strides towards overcoming challenges associated with editing and screening primary human cells.
Extract:
‘CRISPR-Cas9 Knockout Screening Using Primary Human Cells in Drug Discovery’
While significant progress has been made carrying out CRIPR screens in immortalised cell lines, a more physiological and clinically relevant alternative are human primary cells, such as T cells, regulatory B cells, or natural killer (NK) vells. The use of phenotypically relevant primary cells, though, is not without biological and technical challenges, especially when their intended use includes gene editing and scale-up of these specialised cell types for screening. However, overcoming these challenges and utilising new detection technologies for assay readouts herald a new era of cellular screening. Thus, functional genomic screening (FGS) with gene-edited human primary cells offer unique opportunities to accurately identify relevant drug targets and more reliably introduced validated therapeutics to the clinic.
CRISPR-Cas9 Knockout Screening to Understand Gene Function
A logical method to understand the role of a factor within a system is to remove it and assess the result. In drug discovery, this loss-of-function approach is often employed to identify new drug targets by knocking genes down or out and investigating the resulting biological characteristics or phenotype. If the phenotype is unaltered, the target gene likely does not contribute to the phenotype under investigation. However, if the phenotype is altered, then the gene, and its protein, might be a potential drug target. Ideally, these screens involve systematic knockdown or knockout of genes from small scale up to whole genome level and are conducted in a cellular system that resembles in vivo conditions. Harnessing innovative technologies that include gene editing, cell culture, automation, end-point acquisition, and robust analysis is no longer science fiction, but a fully established reality.
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