Using CRISPR/Cas9 Technology for Manipulating Cell Death Regulators.
Clustered, regularly interspaced, short palindromic repeats (CRISPR)/Cas9 technology has been demonstrated to be a useful tool for generating targeted mutations in cell lines and mice. However, the use of CRISPR/Cas9 in a constitutively expressed manner can often result in low targeting efficiencies and lethality due to mutations in essential genes. Here, we describe the use of an inducible lentiviral vector platform, enabling rapid transduction and enrichment of CRISPR/Cas9 positive cells and high levels of targeted mutations upon induction.
Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC, 3052, Australia.|Department of Medical Biology, University of Melbourne, Parkville, VIC, 3050, Australia.
Andrew J
AJ
Kueh
Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC, 3052, Australia. herold@wehi.edu.au.|Department of Medical Biology, University of Melbourne, Parkville, VIC, 3050, Australia. herold@wehi.edu.au.
Marco J
MJ
Herold
D000818:Animals; D064113:CRISPR-Cas Systems; D016923:Cell Death / Q000235:genetics*; D002478:Cells, Cultured; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D017393:RNA Editing
CRISPR/Cas9; Guide RNA; Inducible; Lentiviral