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Polycomb response elements reduce leaky expression of Cas9 under temperature-inducible Hsp70Bb promoter in Drosophila melanogaster

View ORCID ProfileNatalie Warsinger-Pepe, Carly Chang, Connor R. Desroberts, View ORCID ProfileOmar S. Akbari
doi: https://doi.org/10.1101/2022.08.29.505746
Natalie Warsinger-Pepe
1School of Biological Sciences, Department of Cell and Developmental Biology, University of California, San Diego, La Jolla, CA 92093
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Carly Chang
2School of Biological Sciences, University of California, San Diego, La Jolla, CA 92093
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Connor R. Desroberts
2School of Biological Sciences, University of California, San Diego, La Jolla, CA 92093
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Omar S. Akbari
1School of Biological Sciences, Department of Cell and Developmental Biology, University of California, San Diego, La Jolla, CA 92093
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  • For correspondence: oakbari@ucsd.edu
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Abstract

Heat shock inducible expression of genes through the use of heat inducible promoters is commonly used in research despite leaky expression of downstream genes of interest without targeted induction (i.e. heat shock). The development of non-leaky inducible expression systems are of broad interest for both basic and applied studies, to precisely control gene expression. Here we characterize the use of Polycomb response elements and the inducible Heat shock protein 70Bb promoter, previously described as a non-leaky inducible system, to regulate Cas9 endonuclease levels and function in Drosophila melanogaster after varying both heat shock durations and rearing temperatures. We show that Polycomb response elements can significantly reduce expression of Cas9 under Heat shock protein 70Bb promoter control using a range of conditions, corroborating previously published results. We further demonstrate that this low transcript level of heat-induced Cas9 is sufficient to induce mutant mosaic phenotypes. Incomplete suppression of an inducible Cas9 system by Polycomb response elements with no heat shock suggests that further regulatory elements are required to precisely control Cas9 expression and abundance.

Competing Interest Statement

O.S.A is a founder of Agragene, Inc. and Synvect, Inc. with equity interest. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. All other authors declare no competing interests.

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-ND 4.0 International license.
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Posted August 29, 2022.
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Polycomb response elements reduce leaky expression of Cas9 under temperature-inducible Hsp70Bb promoter in Drosophila melanogaster
Natalie Warsinger-Pepe, Carly Chang, Connor R. Desroberts, Omar S. Akbari
bioRxiv 2022.08.29.505746; doi: https://doi.org/10.1101/2022.08.29.505746
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Polycomb response elements reduce leaky expression of Cas9 under temperature-inducible Hsp70Bb promoter in Drosophila melanogaster
Natalie Warsinger-Pepe, Carly Chang, Connor R. Desroberts, Omar S. Akbari
bioRxiv 2022.08.29.505746; doi: https://doi.org/10.1101/2022.08.29.505746

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