An Inducible Lentiviral Guide RNA Platform Enables the Identification of Tumor-Essential Genes and Tumor-Promoting Mutations In Vivo
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
WEHI Research Division(s)
Molecular Genetics Of Cancer; Development And Cancer; Systems Biology And Personalised Medicine
NHMRC Grants
NHMRC/1016701
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
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