A comprehensive protocol resource for performing pooled shRNA and CRISPR screens

Leonie A. Cluse; Iva Nikolic; Deborah Knight; Piyush B. Madhamshettiwar; Jennii Luu; Karla J. Cowley; Timothy Semple; Gisela Mir Arnau; Jake Shortt; Ricky W. Johnstone; Kaylene J. Simpson

doi:10.1007/978-1-4939-7568-6_17

A comprehensive protocol resource for performing pooled shRNA and CRISPR screens

Leonie A. Cluse, Iva Nikolic, Deborah Knight, Piyush B. Madhamshettiwar, Jennii Luu, Karla J. Cowley, Timothy Semple, Gisela Mir Arnau, Jake Shortt, Ricky W. Johnstone, Kaylene J. Simpson

Medicine Monash Health

Research output: Chapter in Book/Report/Conference proceeding › Chapter (Book) › Other › peer-review

3 Citations (Scopus)

Abstract

This chapter details a compendium of protocols that collectively enable the reader to perform a pooled shRNA and/or CRISPR screen—with methods to identify and validate positive controls and subsequent hits; establish a viral titer in the cell line of choice; create and screen libraries, sequence strategies, and bioinformatics resources to analyze outcomes. Collectively, this provides an overarching resource from the start to finish of a screening project, making this technology possible in all laboratories.

Original language	English
Title of host publication	Methods in Molecular Biology
Editors	John M. Walker
Place of Publication	New York, NY
Publisher	Humana Press
Chapter	17
Pages	201-227
Number of pages	27
ISBN (Electronic)	9781493975686
ISBN (Print)	9781493975679
DOIs	https://doi.org/10.1007/978-1-4939-7568-6_17
Publication status	Published - 2018

Publication series

Name	Methods in Molecular Biology
Publisher	Humana Press, Inc.
Volume	1725
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

Drop out screen
Lentivirus
Pooled CRISPR
Pooled shRNA

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10.1007/978-1-4939-7568-6_17

Cite this

Cluse, L. A., Nikolic, I., Knight, D., Madhamshettiwar, P. B., Luu, J., Cowley, K. J., Semple, T., Arnau, G. M., Shortt, J., Johnstone, R. W., & Simpson, K. J. (2018). A comprehensive protocol resource for performing pooled shRNA and CRISPR screens. In J. M. Walker (Ed.), Methods in Molecular Biology (pp. 201-227). (Methods in Molecular Biology; Vol. 1725). Humana Press. https://doi.org/10.1007/978-1-4939-7568-6_17

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title = "A comprehensive protocol resource for performing pooled shRNA and CRISPR screens",

abstract = "This chapter details a compendium of protocols that collectively enable the reader to perform a pooled shRNA and/or CRISPR screen—with methods to identify and validate positive controls and subsequent hits; establish a viral titer in the cell line of choice; create and screen libraries, sequence strategies, and bioinformatics resources to analyze outcomes. Collectively, this provides an overarching resource from the start to finish of a screening project, making this technology possible in all laboratories.",

keywords = "Drop out screen, Lentivirus, Pooled CRISPR, Pooled shRNA",

author = "Cluse, {Leonie A.} and Iva Nikolic and Deborah Knight and Madhamshettiwar, {Piyush B.} and Jennii Luu and Cowley, {Karla J.} and Timothy Semple and Arnau, {Gisela Mir} and Jake Shortt and Johnstone, {Ricky W.} and Simpson, {Kaylene J.}",

year = "2018",

doi = "10.1007/978-1-4939-7568-6_17",

language = "English",

isbn = "9781493975679",

series = "Methods in Molecular Biology",

publisher = "Humana Press",

pages = "201--227",

editor = "Walker, {John M. }",

booktitle = "Methods in Molecular Biology",

address = "United States of America",

}

Cluse, LA, Nikolic, I, Knight, D, Madhamshettiwar, PB, Luu, J, Cowley, KJ, Semple, T, Arnau, GM, Shortt, J, Johnstone, RW & Simpson, KJ 2018, A comprehensive protocol resource for performing pooled shRNA and CRISPR screens. in JM Walker (ed.), Methods in Molecular Biology. Methods in Molecular Biology, vol. 1725, Humana Press, New York, NY, pp. 201-227. https://doi.org/10.1007/978-1-4939-7568-6_17

A comprehensive protocol resource for performing pooled shRNA and CRISPR screens. / Cluse, Leonie A.; Nikolic, Iva; Knight, Deborah et al.
Methods in Molecular Biology. ed. / John M. Walker. New York, NY: Humana Press, 2018. p. 201-227 (Methods in Molecular Biology; Vol. 1725).

Research output: Chapter in Book/Report/Conference proceeding › Chapter (Book) › Other › peer-review

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AU - Cluse, Leonie A.

AU - Nikolic, Iva

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AU - Cowley, Karla J.

AU - Semple, Timothy

AU - Arnau, Gisela Mir

AU - Shortt, Jake

AU - Johnstone, Ricky W.

AU - Simpson, Kaylene J.

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ER -

A comprehensive protocol resource for performing pooled shRNA and CRISPR screens

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Keywords

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