Abstract
Pooled CRISPR screening has emerged as a significant tool within
cancer research. It can be used to interrogate the whole genome to find
potential new drug targets, combination therapies, and resistance mechanisms to
help patient stratification.
Pooled CRISPR screening in primary T cells faces several technical
challenges including efficient Cas9 delivery and sgRNA library transduction.
For this reason, screens are often performed in cell line models, such as
Jurkats, which are known to poorly represent primary T cell biology. We have
adapted the SLICE protocol for genome-wide CRISPR screening in primary T cells
and show that this method is applicable and adaptable across major T cell
sub-populations. We determined that cell age is a key indicator of
transducibility allowing us to forecast viral requirements based on cell age. We
have evolved this method further to now be able to integrate biologically
relevant genetic modifications into the process, in order to utilise this
platform to support the target/drug discovery and development process across
the AZ and CRUK communities.
