Combinatorial CRISPR-Cas9 Knockout Screen
Simultaneous mutation of two genes can produce a phenotype that is unexpected in light of each mutation’s individual effect. This phenomenon, known as genetic interaction, identifies an underlying functional relationship between the genes, such as contributions to the same protein complex or pathway. Mapping these functional relationships has broad applicability for advancing our fundamental understanding of biological systems and also has direct implications for cancer therapeutic development. Here we describe a method using the CRISPR-Cas9 system to knockout pairs of genes, enabling high-throughput, systematic mapping of these genetic interaction networks in human cells. This protocol accompanies Shen et al (Nature Methods, publishing online 20 March, 2017; 10.038/nmeth.4225); it was added to the manuscript after formal peer review, as an aid to users.
Figure 1
Dear Dr. Mali, did you integrate Cas9 into AAVS1? Did you test Cas9 activity? THX.
Posted 20 Mar, 2017
Combinatorial CRISPR-Cas9 Knockout Screen
Posted 20 Mar, 2017
Simultaneous mutation of two genes can produce a phenotype that is unexpected in light of each mutation’s individual effect. This phenomenon, known as genetic interaction, identifies an underlying functional relationship between the genes, such as contributions to the same protein complex or pathway. Mapping these functional relationships has broad applicability for advancing our fundamental understanding of biological systems and also has direct implications for cancer therapeutic development. Here we describe a method using the CRISPR-Cas9 system to knockout pairs of genes, enabling high-throughput, systematic mapping of these genetic interaction networks in human cells. This protocol accompanies Shen et al (Nature Methods, publishing online 20 March, 2017; 10.038/nmeth.4225); it was added to the manuscript after formal peer review, as an aid to users.
Figure 1
Dear Dr. Mali, did you integrate Cas9 into AAVS1? Did you test Cas9 activity? THX.
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