CRISPR knockout screening outperforms shRNA and CRISPRi in identifying essential genes.
High-throughput genetic screens have become essential tools for studying a wide variety of biological processes. Here we experimentally compare systems based on clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) or its transcriptionally repressive variant, CRISPR-interference (CRISPRi), with a traditional short hairpin RNA (shRNA)-based system for performing lethality screens. We find that the CRISPR technology performed best, with low noise, minimal off-target effects and consistent activity across reagents.
Division of Molecular Carcinogenesis, Cancer Systems Biology Centre and Cancer Genomics Centre Netherlands, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
Bastiaan
B
Evers
Division of Molecular Carcinogenesis, Cancer Systems Biology Centre and Cancer Genomics Centre Netherlands, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
Katarzyna
K
Jastrzebski
Division of Molecular Carcinogenesis, Cancer Systems Biology Centre and Cancer Genomics Centre Netherlands, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
Jeroen P M
JP
Heijmans
Division of Molecular Carcinogenesis, Cancer Systems Biology Centre and Cancer Genomics Centre Netherlands, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
Wipawadee
W
Grernrum
Division of Molecular Carcinogenesis, Cancer Systems Biology Centre and Cancer Genomics Centre Netherlands, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
Roderick L
RL
Beijersbergen
Division of Molecular Carcinogenesis, Cancer Systems Biology Centre and Cancer Genomics Centre Netherlands, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
Rene
http://orcid.org/0000-0001-8677-3423
R
Bernards
D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods*; D055786:Gene Knockout Techniques / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods*; D034741:RNA, Small Interfering / Q000235:genetics*
D005819:Genetic Markers; D034741:RNA, Small Interfering