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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2021-03-01
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Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721000289 |
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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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1724207565033701376 |