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A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen.

Publication ,  Journal Article
Moffat, J; Grueneberg, DA; Yang, X; Kim, SY; Kloepfer, AM; Hinkle, G; Piqani, B; Eisenhaure, TM; Luo, B; Grenier, JK; Carpenter, AE; Foo, SY ...
Published in: Cell
March 24, 2006

To enable arrayed or pooled loss-of-function screens in a wide range of mammalian cell types, including primary and nondividing cells, we are developing lentiviral short hairpin RNA (shRNA) libraries targeting the human and murine genomes. The libraries currently contain 104,000 vectors, targeting each of 22,000 human and mouse genes with multiple sequence-verified constructs. To test the utility of the library for arrayed screens, we developed a screen based on high-content imaging to identify genes required for mitotic progression in human cancer cells and applied it to an arrayed set of 5,000 unique shRNA-expressing lentiviruses that target 1,028 human genes. The screen identified several known and approximately 100 candidate regulators of mitotic progression and proliferation; the availability of multiple shRNAs targeting the same gene facilitated functional validation of putative hits. This work provides a widely applicable resource for loss-of-function screens, as well as a roadmap for its application to biological discovery.

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Published In

Cell

DOI

ISSN

0092-8674

Publication Date

March 24, 2006

Volume

124

Issue

6

Start / End Page

1283 / 1298

Location

United States

Related Subject Headings

  • RNA, Small Interfering
  • Microarray Analysis
  • Mice
  • Libraries
  • Lentivirus
  • Humans
  • Genetic Vectors
  • Genetic Engineering
  • Gene Library
  • Developmental Biology
 

Citation

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Moffat, J., Grueneberg, D. A., Yang, X., Kim, S. Y., Kloepfer, A. M., Hinkle, G., … Root, D. E. (2006). A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen. Cell, 124(6), 1283–1298. https://doi.org/10.1016/j.cell.2006.01.040
Moffat, Jason, Dorre A. Grueneberg, Xiaoping Yang, So Young Kim, Angela M. Kloepfer, Gregory Hinkle, Bruno Piqani, et al. “A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen.Cell 124, no. 6 (March 24, 2006): 1283–98. https://doi.org/10.1016/j.cell.2006.01.040.
Moffat J, Grueneberg DA, Yang X, Kim SY, Kloepfer AM, Hinkle G, et al. A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen. Cell. 2006 Mar 24;124(6):1283–98.
Moffat, Jason, et al. “A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen.Cell, vol. 124, no. 6, Mar. 2006, pp. 1283–98. Pubmed, doi:10.1016/j.cell.2006.01.040.
Moffat J, Grueneberg DA, Yang X, Kim SY, Kloepfer AM, Hinkle G, Piqani B, Eisenhaure TM, Luo B, Grenier JK, Carpenter AE, Foo SY, Stewart SA, Stockwell BR, Hacohen N, Hahn WC, Lander ES, Sabatini DM, Root DE. A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen. Cell. 2006 Mar 24;124(6):1283–1298.
Journal cover image

Published In

Cell

DOI

ISSN

0092-8674

Publication Date

March 24, 2006

Volume

124

Issue

6

Start / End Page

1283 / 1298

Location

United States

Related Subject Headings

  • RNA, Small Interfering
  • Microarray Analysis
  • Mice
  • Libraries
  • Lentivirus
  • Humans
  • Genetic Vectors
  • Genetic Engineering
  • Gene Library
  • Developmental Biology