Journal
CELL REPORTS
Volume 10, Issue 8, Pages 1422-1432Publisher
CELL PRESS
DOI: 10.1016/j.celrep.2015.02.002
Keywords
-
Categories
Funding
- Leukemia Foundation National Research Program Clinical PhD Scholarship
- Kay Kendall Leukemia Fund Intermediate Fellowship [KKL331]
- National Health and Medical Research Council, Australia program [1016701, 1020363]
- Leukemia and Lymphoma Society SCOR [7001-13, GNT1049720]
Ask authors/readers for more resources
The CRISPR/Cas9 technology enables the introduction of genomic alterations into almost any organism; however, systems for efficient and inducible gene modification have been lacking, especially for deletion of essential genes. Here, we describe a drug-inducible small guide RNA (sgRNA) vector system allowing for ubiquitous and efficient gene deletion in murine and human cells. This system mediates the efficient, temporally controlled deletion of MCL-1, both in vitro and in vivo, in human Burkitt lymphoma cell lines that require this anti-apoptotic BCL-2 protein for sustained survival and growth. Unexpectedly, repeated induction of the same sgRNA generated similar inactivating mutations in the human Mcl-1 gene due to low mutation variability exerted by the accompanying non-homologous end-joining (NHEJ) process. Finally, we were able to generate hematopoietic cell compartment-restricted Trp53-knockout mice, leading to the identification of cancer-promoting mutants of this critical tumor suppressor.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available