Cargando…
Considerations and practical implications of performing a phenotypic CRISPR/Cas survival screen
Genome-wide screens that have viability as a readout have been instrumental to identify essential genes. The development of gene knockout screens with the use of CRISPR-Cas has provided a more sensitive method to identify these genes. Here, we performed an exhaustive genome-wide CRISPR/Cas9 phenotyp...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8853573/ https://www.ncbi.nlm.nih.gov/pubmed/35176052 http://dx.doi.org/10.1371/journal.pone.0263262 |
_version_ | 1784653260593823744 |
---|---|
author | Ashoti, Ator Limone, Francesco van Kranenburg, Melissa Alemany, Anna Baak, Mirna Vivié, Judith Piccioni, Frederica Dijkers, Pascale F. Creyghton, Menno Eggan, Kevin Geijsen, Niels |
author_facet | Ashoti, Ator Limone, Francesco van Kranenburg, Melissa Alemany, Anna Baak, Mirna Vivié, Judith Piccioni, Frederica Dijkers, Pascale F. Creyghton, Menno Eggan, Kevin Geijsen, Niels |
author_sort | Ashoti, Ator |
collection | PubMed |
description | Genome-wide screens that have viability as a readout have been instrumental to identify essential genes. The development of gene knockout screens with the use of CRISPR-Cas has provided a more sensitive method to identify these genes. Here, we performed an exhaustive genome-wide CRISPR/Cas9 phenotypic rescue screen to identify modulators of cytotoxicity induced by the pioneer transcription factor, DUX4. Misexpression of DUX4 due to a failure in epigenetic repressive mechanisms underlies facioscapulohumeral muscular dystrophy (FHSD), a complex muscle disorder that thus far remains untreatable. As the name implies, FSHD generally starts in the muscles of the face and shoulder girdle. Our CRISPR/Cas9 screen revealed no key effectors other than DUX4 itself that could modulate DUX4 cytotoxicity, suggesting that treatment efforts in FSHD should be directed towards direct modulation of DUX4 itself. Our screen did however reveal some rare and unexpected genomic events, that had an important impact on the interpretation of our data. Our findings may provide important considerations for planning future CRISPR/Cas9 phenotypic survival screens. |
format | Online Article Text |
id | pubmed-8853573 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-88535732022-02-18 Considerations and practical implications of performing a phenotypic CRISPR/Cas survival screen Ashoti, Ator Limone, Francesco van Kranenburg, Melissa Alemany, Anna Baak, Mirna Vivié, Judith Piccioni, Frederica Dijkers, Pascale F. Creyghton, Menno Eggan, Kevin Geijsen, Niels PLoS One Research Article Genome-wide screens that have viability as a readout have been instrumental to identify essential genes. The development of gene knockout screens with the use of CRISPR-Cas has provided a more sensitive method to identify these genes. Here, we performed an exhaustive genome-wide CRISPR/Cas9 phenotypic rescue screen to identify modulators of cytotoxicity induced by the pioneer transcription factor, DUX4. Misexpression of DUX4 due to a failure in epigenetic repressive mechanisms underlies facioscapulohumeral muscular dystrophy (FHSD), a complex muscle disorder that thus far remains untreatable. As the name implies, FSHD generally starts in the muscles of the face and shoulder girdle. Our CRISPR/Cas9 screen revealed no key effectors other than DUX4 itself that could modulate DUX4 cytotoxicity, suggesting that treatment efforts in FSHD should be directed towards direct modulation of DUX4 itself. Our screen did however reveal some rare and unexpected genomic events, that had an important impact on the interpretation of our data. Our findings may provide important considerations for planning future CRISPR/Cas9 phenotypic survival screens. Public Library of Science 2022-02-17 /pmc/articles/PMC8853573/ /pubmed/35176052 http://dx.doi.org/10.1371/journal.pone.0263262 Text en © 2022 Ashoti et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Ashoti, Ator Limone, Francesco van Kranenburg, Melissa Alemany, Anna Baak, Mirna Vivié, Judith Piccioni, Frederica Dijkers, Pascale F. Creyghton, Menno Eggan, Kevin Geijsen, Niels Considerations and practical implications of performing a phenotypic CRISPR/Cas survival screen |
title | Considerations and practical implications of performing a phenotypic CRISPR/Cas survival screen |
title_full | Considerations and practical implications of performing a phenotypic CRISPR/Cas survival screen |
title_fullStr | Considerations and practical implications of performing a phenotypic CRISPR/Cas survival screen |
title_full_unstemmed | Considerations and practical implications of performing a phenotypic CRISPR/Cas survival screen |
title_short | Considerations and practical implications of performing a phenotypic CRISPR/Cas survival screen |
title_sort | considerations and practical implications of performing a phenotypic crispr/cas survival screen |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8853573/ https://www.ncbi.nlm.nih.gov/pubmed/35176052 http://dx.doi.org/10.1371/journal.pone.0263262 |
work_keys_str_mv | AT ashotiator considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT limonefrancesco considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT vankranenburgmelissa considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT alemanyanna considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT baakmirna considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT viviejudith considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT piccionifrederica considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT dijkerspascalef considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT creyghtonmenno considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT eggankevin considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen AT geijsenniels considerationsandpracticalimplicationsofperformingaphenotypiccrisprcassurvivalscreen |