A rapid and scalable system for studying gene function in mice using conditional RNA interference

Prem K. Premsrirut; Lukas E. Dow; Sang Yong Kim; Matthew Camiolo; Colin D. Malone; Cornelius Miething; Claudio Scuoppo; Johannes Zuber; Ross A. Dickins; Scott C. Kogan; Kenneth R. Shroyer; Raffaella Sordella; Gregory J. Hannon; Scott W. Lowe

doi:10.1016/j.cell.2011.03.012

A rapid and scalable system for studying gene function in mice using conditional RNA interference

Prem K. Premsrirut
, Lukas E. Dow
, Sang Yong Kim
, Matthew Camiolo
, Colin D. Malone
, Cornelius Miething
, Claudio Scuoppo
, Johannes Zuber
, Ross A. Dickins
, Scott C. Kogan
, Kenneth R. Shroyer
, Raffaella Sordella
, Gregory J. Hannon
, Scott W. Lowe

Pathology

Cold Spring Harbor Laboratory
Stony Brook University
Watson School of Biological Sciences
Walter and Eliza Hall Institute of Medical Research
University of California at San Francisco
Howard Hughes Medical Institute

Research output: Contribution to journal › Article › peer-review

266 Scopus citations

Abstract

RNA interference is a powerful tool for studying gene function, however, the reproducible generation of RNAi transgenic mice remains a significant limitation. By combining optimized fluorescence-coupled miR30-based shRNAs with high efficiency ES cell targeting, we developed a fast, scalable pipeline for the production of shRNA transgenic mice. Using this system, we generated eight tet-regulated shRNA transgenic lines targeting Firefly and Renilla luciferases, Oct4 and tumor suppressors p53, p16^INK4a, p19^ARF and APC and demonstrate potent gene silencing and GFP-tracked knockdown in a broad range of tissues in vivo. Further, using an shRNA targeting APC, we illustrate how this approach can identify predicted phenotypes and also unknown functions for a well-studied gene. In addition, through regulated gene silencing we validate APC/Wnt and p19^ARF as potential therapeutic targets in T cell acute lymphoblastic leukemia/lymphoma and lung adenocarcinoma, respectively. This system provides a cost-effective and scalable platform for the production of RNAi transgenic mice targeting any mammalian gene. PaperClip:

Original language	English
Pages (from-to)	145-158
Number of pages	14
Journal	Cell
Volume	145
Issue number	1
DOIs	https://doi.org/10.1016/j.cell.2011.03.012
State	Published - Apr 1 2011

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10.1016/j.cell.2011.03.012

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Premsrirut, P. K., Dow, L. E., Kim, S. Y., Camiolo, M., Malone, C. D., Miething, C., Scuoppo, C., Zuber, J., Dickins, R. A., Kogan, S. C., Shroyer, K. R., Sordella, R., Hannon, G. J., & Lowe, S. W. (2011). A rapid and scalable system for studying gene function in mice using conditional RNA interference. Cell, 145(1), 145-158. https://doi.org/10.1016/j.cell.2011.03.012

@article{b8bffc59235c492dabd8a394adac00be,

title = "A rapid and scalable system for studying gene function in mice using conditional RNA interference",

abstract = "RNA interference is a powerful tool for studying gene function, however, the reproducible generation of RNAi transgenic mice remains a significant limitation. By combining optimized fluorescence-coupled miR30-based shRNAs with high efficiency ES cell targeting, we developed a fast, scalable pipeline for the production of shRNA transgenic mice. Using this system, we generated eight tet-regulated shRNA transgenic lines targeting Firefly and Renilla luciferases, Oct4 and tumor suppressors p53, p16INK4a, p19ARF and APC and demonstrate potent gene silencing and GFP-tracked knockdown in a broad range of tissues in vivo. Further, using an shRNA targeting APC, we illustrate how this approach can identify predicted phenotypes and also unknown functions for a well-studied gene. In addition, through regulated gene silencing we validate APC/Wnt and p19ARF as potential therapeutic targets in T cell acute lymphoblastic leukemia/lymphoma and lung adenocarcinoma, respectively. This system provides a cost-effective and scalable platform for the production of RNAi transgenic mice targeting any mammalian gene. PaperClip:",

author = "Premsrirut, \{Prem K.\} and Dow, \{Lukas E.\} and Kim, \{Sang Yong\} and Matthew Camiolo and Malone, \{Colin D.\} and Cornelius Miething and Claudio Scuoppo and Johannes Zuber and Dickins, \{Ross A.\} and Kogan, \{Scott C.\} and Shroyer, \{Kenneth R.\} and Raffaella Sordella and Hannon, \{Gregory J.\} and Lowe, \{Scott W.\}",

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AU - Premsrirut, Prem K.

AU - Dow, Lukas E.

AU - Kim, Sang Yong

AU - Camiolo, Matthew

AU - Malone, Colin D.

AU - Miething, Cornelius

AU - Scuoppo, Claudio

AU - Zuber, Johannes

AU - Dickins, Ross A.

AU - Kogan, Scott C.

AU - Shroyer, Kenneth R.

AU - Sordella, Raffaella

AU - Hannon, Gregory J.

AU - Lowe, Scott W.

PY - 2011/4/1

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N2 - RNA interference is a powerful tool for studying gene function, however, the reproducible generation of RNAi transgenic mice remains a significant limitation. By combining optimized fluorescence-coupled miR30-based shRNAs with high efficiency ES cell targeting, we developed a fast, scalable pipeline for the production of shRNA transgenic mice. Using this system, we generated eight tet-regulated shRNA transgenic lines targeting Firefly and Renilla luciferases, Oct4 and tumor suppressors p53, p16INK4a, p19ARF and APC and demonstrate potent gene silencing and GFP-tracked knockdown in a broad range of tissues in vivo. Further, using an shRNA targeting APC, we illustrate how this approach can identify predicted phenotypes and also unknown functions for a well-studied gene. In addition, through regulated gene silencing we validate APC/Wnt and p19ARF as potential therapeutic targets in T cell acute lymphoblastic leukemia/lymphoma and lung adenocarcinoma, respectively. This system provides a cost-effective and scalable platform for the production of RNAi transgenic mice targeting any mammalian gene. PaperClip:

AB - RNA interference is a powerful tool for studying gene function, however, the reproducible generation of RNAi transgenic mice remains a significant limitation. By combining optimized fluorescence-coupled miR30-based shRNAs with high efficiency ES cell targeting, we developed a fast, scalable pipeline for the production of shRNA transgenic mice. Using this system, we generated eight tet-regulated shRNA transgenic lines targeting Firefly and Renilla luciferases, Oct4 and tumor suppressors p53, p16INK4a, p19ARF and APC and demonstrate potent gene silencing and GFP-tracked knockdown in a broad range of tissues in vivo. Further, using an shRNA targeting APC, we illustrate how this approach can identify predicted phenotypes and also unknown functions for a well-studied gene. In addition, through regulated gene silencing we validate APC/Wnt and p19ARF as potential therapeutic targets in T cell acute lymphoblastic leukemia/lymphoma and lung adenocarcinoma, respectively. This system provides a cost-effective and scalable platform for the production of RNAi transgenic mice targeting any mammalian gene. PaperClip:

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VL - 145

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EP - 158

JO - Cell

JF - Cell

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ER -

A rapid and scalable system for studying gene function in mice using conditional RNA interference

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