Reversible suppression of an essential gene in adult mice using transgenic RNA interference

McJunkin, K., Mazurek, A., Premsrirut, P. K., Zuber, J., Dow, L. E., Simon, J., Stillman, B., Lowe, S. W. (2011) Reversible suppression of an essential gene in adult mice using transgenic RNA interference. Proceedings of the National Academy of Sciences of the United States of America, 108 (17). pp. 7113-7118. ISSN 0027-8424

URL: http://www.ncbi.nlm.nih.gov/pubmed/21482754
DOI: 10.1073/pnas.1104097108

Abstract

RNAi has revolutionized loss-of-function genetics by enabling sequence-specific suppression of virtually any gene. Furthermore, tetracycline response elements (TRE) can drive expression of short hairpin RNAs (shRNAs) for inducible and reversible target gene suppression. Here, we demonstrate the feasibility of transgenic inducible RNAi for suppression of essential genes. We set out to directly target cell proliferation by screening an RNAi library against DNA replication factors and identified multiple shRNAs against Replication Protein A, subunit 3 (RPA3). We generated transgenic mice with TRE-driven Rpa3 shRNAs whose expression enforced a reversible cell cycle arrest. In adult mice, the block in cell proliferation caused rapid atrophy of the intestinal epithelium which led to weight loss and lethality within 8-11 d of shRNA induction. Upon shRNA withdrawal, villus atrophy and weight loss were fully reversible. Thus, shRpa3 transgenic mice provide an interesting tool to study tissue maintenance and regeneration. Overall, we have established a robust system that serves the purpose of temperature-sensitive alleles in other model organisms, enabling inducible and reversible suppression of essential genes in a mammalian system.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNAi
CSHL Authors:
Communities: CSHL Cancer Center Program > Cancer Genetics
CSHL Cancer Center Program > Gene Regulation and Cell Proliferation
CSHL Cancer Center Shared Resources > Animal Services
CSHL Cancer Center Shared Resources > DNA Sequencing Service
CSHL Cancer Center Shared Resources > Flow Cytometry Service
CSHL Cancer Center Shared Resources > Gene Targeting Service
CSHL Post Doctoral Fellows
CSHL labs > Lowe lab
CSHL labs > Stillman lab
Watson School > Publications
CSHL Cancer Center Shared Resources > Functional Genomics and Genetics Service
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date Deposited: 06 Mar 2012 14:42
Last Modified: 20 Jun 2017 16:29
PMCID: PMC3084121
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25017

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