Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library

Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library

Summary: CRISPR-based genetic screens revolutionized our ability to genetically probe cell biology. We present a protocol to conduct genome-scale chemogenomic dropout CRISPR screens in the human RPE1-hTERT p53−/− cell line. We use the TKOv3 library, which contains 70,948 sgRNAs targeting 18,053 gene...

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Main Authors: Michele Olivieri, Daniel Durocher
Format: Article
Language:English
Published: Elsevier 2021-03-01
Series:STAR Protocols
Subjects:
CRISPR
Genetics
Genomics
High-throughput screening
Sequence analysis
Sequencing
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721000289
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spelling doaj-6f61fec1e3884144b21c9e32904434552021-03-22T12:53:13ZengElsevierSTAR Protocols2666-16672021-03-0121100321Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 libraryMichele Olivieri0Daniel Durocher1Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, ON M5G 1X5, Canada; Department of Molecular Genetics, University of Toronto, 1 King’s College Circle, Toronto, ON M5S 1A8, CanadaLunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, ON M5G 1X5, Canada; Department of Molecular Genetics, University of Toronto, 1 King’s College Circle, Toronto, ON M5S 1A8, Canada; Corresponding authorSummary: CRISPR-based genetic screens revolutionized our ability to genetically probe cell biology. We present a protocol to conduct genome-scale chemogenomic dropout CRISPR screens in the human RPE1-hTERT p53−/− cell line. We use the TKOv3 library, which contains 70,948 sgRNAs targeting 18,053 genes. Here, we describe how to set up the screen, the reagents required, and how to sequence and analyze the results. This protocol can be customized for other libraries, cell lines, and sequencing instruments.For complete details on the use and execution of this protocol, please refer to Olivieri et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166721000289CRISPRGeneticsGenomicsHigh-throughput screeningSequence analysisSequencing
collection DOAJ
language English
format Article
sources DOAJ
author Michele Olivieri
Daniel Durocher
spellingShingle Michele Olivieri
Daniel Durocher
Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library
STAR Protocols
CRISPR
Genetics
Genomics
High-throughput screening
Sequence analysis
Sequencing
author_facet Michele Olivieri
Daniel Durocher
author_sort Michele Olivieri
title Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library
title_short Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library
title_full Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library
title_fullStr Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library
title_full_unstemmed Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library
title_sort genome-scale chemogenomic crispr screens in human cells using the tkov3 library
publisher Elsevier
series STAR Protocols
issn 2666-1667
publishDate 2021-03-01
description Summary: CRISPR-based genetic screens revolutionized our ability to genetically probe cell biology. We present a protocol to conduct genome-scale chemogenomic dropout CRISPR screens in the human RPE1-hTERT p53−/− cell line. We use the TKOv3 library, which contains 70,948 sgRNAs targeting 18,053 genes. Here, we describe how to set up the screen, the reagents required, and how to sequence and analyze the results. This protocol can be customized for other libraries, cell lines, and sequencing instruments.For complete details on the use and execution of this protocol, please refer to Olivieri et al. (2020).
topic CRISPR
Genetics
Genomics
High-throughput screening
Sequence analysis
Sequencing
url http://www.sciencedirect.com/science/article/pii/S2666166721000289
work_keys_str_mv AT micheleolivieri genomescalechemogenomiccrisprscreensinhumancellsusingthetkov3library
AT danieldurocher genomescalechemogenomiccrisprscreensinhumancellsusingthetkov3library
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