A CRISPR RESOURCE FOR INDIVIDUAL, COMBINATORIAL OR MULTIPLEXED GENE KNOCKOUT
CRISPR is a new technology for genetic engineering of human (and many other) cells. This study created a resource for efficiently knocking out 1 or more genes in cell culture so their effects can be studied in a dish.
We have combined a machine-learning approach with other strategies to optimize knockout efficiency with the CRISPR/Cas9 system. In addition, we have developed a multiplexed sgRNA expression strategy that promotes the functional ablation of single genes and allows for combinatorial targeting. These strategies have been combined to design and construct a genome-wide, sequence-verified, arrayed CRISPR library. This resource allows single-target or combinatorial genetic screens to be carried out at scale in a multiplexed or arrayed format. By conducting parallel loss-of-function screens, we compare our approach to existing sgRNA design and expression strategies.
RELEVANCE TO OC
This is one of the primary methods we use today to study ovarian cancer driver and risk genes, including their associated regulatory elements.