A cellular stress response induced by the CRISPR/dCas9 activation system is not heritable through cell divisions

Andrew D. Johnston; Alali Abdulrazak; Hanae Sato; Shahina B. Maqbool; Masako Suzuki; John M. Greally; Claudia A. Simões-Pires

doi:10.1101/2019.12.23.887224

A cellular stress response induced by the CRISPR/dCas9 activation system is not heritable through cell divisions

Andrew D. Johnston, Alali Abdulrazak, Hanae Sato, Shahina B. Maqbool, Masako Suzuki, John M. Greally, Claudia A. Simões-Pires

Research output: Contribution to journal › Article › peer-review

Abstract

The CRISPR/Cas9 system can be modified to perform ‘epigenetic editing’ by utilizing the catalytically-inactive (dead) Cas9 (dCas9) to recruit regulatory proteins to specific genomic locations. In prior studies, epigenetic editing with multimers of the transactivator VP16 and guide RNAs (gRNAs) was found to cause adverse cellular responses. These side effects may confound studies inducing new cellular properties, especially if the cellular responses are maintained through cell divisions - an epigenetic regulatory property. Here we show how distinct components of this CRISPR/dCas9 activation system, particularly untargeted gRNAs, upregulate genes associated with transcriptional stress, defense response, and regulation of cell death. Our results highlight a previously undetected acute stress response to CRISPR/dCas9 components in human cells, which is transient and not maintained through cell divisions.

Original language	English (US)
Journal	Unknown Journal
DOIs	https://doi.org/10.1101/2019.12.23.887224
State	Published - Dec 23 2019

Keywords

cell division
CRISPR
guide RNA
stress response genes
VP16

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Agricultural and Biological Sciences(all)
Immunology and Microbiology(all)
Neuroscience(all)
Pharmacology, Toxicology and Pharmaceutics(all)

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10.1101/2019.12.23.887224

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title = "A cellular stress response induced by the CRISPR/dCas9 activation system is not heritable through cell divisions",

abstract = "The CRISPR/Cas9 system can be modified to perform {\textquoteleft}epigenetic editing{\textquoteright} by utilizing the catalytically-inactive (dead) Cas9 (dCas9) to recruit regulatory proteins to specific genomic locations. In prior studies, epigenetic editing with multimers of the transactivator VP16 and guide RNAs (gRNAs) was found to cause adverse cellular responses. These side effects may confound studies inducing new cellular properties, especially if the cellular responses are maintained through cell divisions - an epigenetic regulatory property. Here we show how distinct components of this CRISPR/dCas9 activation system, particularly untargeted gRNAs, upregulate genes associated with transcriptional stress, defense response, and regulation of cell death. Our results highlight a previously undetected acute stress response to CRISPR/dCas9 components in human cells, which is transient and not maintained through cell divisions.",

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author = "Johnston, {Andrew D.} and Alali Abdulrazak and Hanae Sato and Maqbool, {Shahina B.} and Masako Suzuki and Greally, {John M.} and Sim{\~o}es-Pires, {Claudia A.}",

note = "Publisher Copyright: The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

year = "2019",

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AU - Abdulrazak, Alali

AU - Sato, Hanae

AU - Maqbool, Shahina B.

AU - Suzuki, Masako

AU - Greally, John M.

AU - Simões-Pires, Claudia A.

N1 - Publisher Copyright: The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2019/12/23

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N2 - The CRISPR/Cas9 system can be modified to perform ‘epigenetic editing’ by utilizing the catalytically-inactive (dead) Cas9 (dCas9) to recruit regulatory proteins to specific genomic locations. In prior studies, epigenetic editing with multimers of the transactivator VP16 and guide RNAs (gRNAs) was found to cause adverse cellular responses. These side effects may confound studies inducing new cellular properties, especially if the cellular responses are maintained through cell divisions - an epigenetic regulatory property. Here we show how distinct components of this CRISPR/dCas9 activation system, particularly untargeted gRNAs, upregulate genes associated with transcriptional stress, defense response, and regulation of cell death. Our results highlight a previously undetected acute stress response to CRISPR/dCas9 components in human cells, which is transient and not maintained through cell divisions.

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