Open AccessConsiderations and practical implications of performing a phenotypic CRISPR/Cas survival screen
Author(s) -
Ator Ashoti,
Francesco Limone,
Melissa van Kranenburg,
Anna Alemany,
Mirna Baak,
Judith Vivié,
Federica Piccioni,
Pascale F. Dijkers,
Menno P. Creyghton,
Kevin Eggan,
Niels Geijsen
Publication year - 2022
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0263262
Subject(s) - crispr , facioscapulohumeral muscular dystrophy , biology , genome editing , cas9 , epigenetics , phenotype , effector , genetics , computational biology , gene , transcription factor , microbiology and biotechnology
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.