An Inducible Lentiviral Guide RNA Platform Enables the Identification of Tumor-Essential Genes and Tumor-Promoting Mutations In Vivo
- Author(s)
- Aubrey, BJ; Kelly, GL; Kueh, AJ; Brennan, MS; O'Connor, L; Milla, L; Wilcox, S; Tai, L; Strasser, A; Herold, MJ;
- Details
- Publication Year 2015-02-24,Volume 10,Issue #8,Page 1422-1432
- Journal Title
- Cell Rep
- Publication Type
- Journal Article
- Abstract
- 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.
- Publisher
- Cell Press
- Research Division(s)
- Molecular Genetics Of Cancer; Development And Cancer; Systems Biology And Personalised Medicine
- Publisher's Version
- https://doi.org/10.1016/j.celrep.2015.02.002
- Open Access at Publisher's Site
http://www.sciencedirect.com/science/article/pii/S2211124715001102- NHMRC Grants
- NHMRC/1016701,
- Terms of Use/Rights Notice
- Copyright © 2015 The Authors. Published by Elsevier Inc.This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).
Creation Date: 2015-03-05 08:08:48
Last Modified: 2015-05-12 09:12:52