Non-Mendelian Dominant Maternal Effects Caused by CRISPR/Cas9 Transgenic Components in Drosophila melanogaster

The CRISPR/Cas9 system has revolutionized genomic editing. The Cas9 endonuclease targets DNA via an experimentally determined guide RNA (gRNA). This results in a double-strand break at the target site . We generated transgenic Drosophila melanogaster in which the CRISPR/Cas9 system was used to targe...

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Main Authors: Chun-Chieh Lin, Christopher J. Potter
Format: Article
Language:English
Published: Oxford University Press 2016-11-01
Series:G3: Genes, Genomes, Genetics
Subjects:
MCR
Online Access:http://g3journal.org/lookup/doi/10.1534/g3.116.034884
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spelling doaj-2142d8479e7946a8aa57a124bfb08b712021-07-02T18:13:48ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362016-11-016113685369110.1534/g3.116.03488425Non-Mendelian Dominant Maternal Effects Caused by CRISPR/Cas9 Transgenic Components in Drosophila melanogasterChun-Chieh LinChristopher J. PotterThe CRISPR/Cas9 system has revolutionized genomic editing. The Cas9 endonuclease targets DNA via an experimentally determined guide RNA (gRNA). This results in a double-strand break at the target site . We generated transgenic Drosophila melanogaster in which the CRISPR/Cas9 system was used to target a GAL4 transgene in vivo. To our surprise, progeny whose genomes did not contain CRISPR/Cas9 components were still capable of mutating GAL4 sequences. We demonstrate this effect was caused by maternal deposition of Cas9 and gRNAs into the embryo, leading to extensive GAL4 mutations in both somatic and germline tissues. This serves as a cautionary observation on the effects of maternal contributions when conducting experiments using genomically encoded CRISPR/Cas9 components. These results also highlight a mode of artificial inheritance in which maternal contributions of DNA editing components lead to transmissible mutant defects even in animals whose genomes lack the editing components. We suggest calling this a dominant maternal effect to reflect it is caused by the gain of maternally contributed products. Models of CRISPR-mediated gene drive will need to incorporate dominant maternal effects in order to accurately predict the efficiency and dynamics of gene drive in a population.http://g3journal.org/lookup/doi/10.1534/g3.116.034884gene drivematernal effectHACKmutagenic chain reactionMCR
collection DOAJ
language English
format Article
sources DOAJ
author Chun-Chieh Lin
Christopher J. Potter
spellingShingle Chun-Chieh Lin
Christopher J. Potter
Non-Mendelian Dominant Maternal Effects Caused by CRISPR/Cas9 Transgenic Components in Drosophila melanogaster
G3: Genes, Genomes, Genetics
gene drive
maternal effect
HACK
mutagenic chain reaction
MCR
author_facet Chun-Chieh Lin
Christopher J. Potter
author_sort Chun-Chieh Lin
title Non-Mendelian Dominant Maternal Effects Caused by CRISPR/Cas9 Transgenic Components in Drosophila melanogaster
title_short Non-Mendelian Dominant Maternal Effects Caused by CRISPR/Cas9 Transgenic Components in Drosophila melanogaster
title_full Non-Mendelian Dominant Maternal Effects Caused by CRISPR/Cas9 Transgenic Components in Drosophila melanogaster
title_fullStr Non-Mendelian Dominant Maternal Effects Caused by CRISPR/Cas9 Transgenic Components in Drosophila melanogaster
title_full_unstemmed Non-Mendelian Dominant Maternal Effects Caused by CRISPR/Cas9 Transgenic Components in Drosophila melanogaster
title_sort non-mendelian dominant maternal effects caused by crispr/cas9 transgenic components in drosophila melanogaster
publisher Oxford University Press
series G3: Genes, Genomes, Genetics
issn 2160-1836
publishDate 2016-11-01
description The CRISPR/Cas9 system has revolutionized genomic editing. The Cas9 endonuclease targets DNA via an experimentally determined guide RNA (gRNA). This results in a double-strand break at the target site . We generated transgenic Drosophila melanogaster in which the CRISPR/Cas9 system was used to target a GAL4 transgene in vivo. To our surprise, progeny whose genomes did not contain CRISPR/Cas9 components were still capable of mutating GAL4 sequences. We demonstrate this effect was caused by maternal deposition of Cas9 and gRNAs into the embryo, leading to extensive GAL4 mutations in both somatic and germline tissues. This serves as a cautionary observation on the effects of maternal contributions when conducting experiments using genomically encoded CRISPR/Cas9 components. These results also highlight a mode of artificial inheritance in which maternal contributions of DNA editing components lead to transmissible mutant defects even in animals whose genomes lack the editing components. We suggest calling this a dominant maternal effect to reflect it is caused by the gain of maternally contributed products. Models of CRISPR-mediated gene drive will need to incorporate dominant maternal effects in order to accurately predict the efficiency and dynamics of gene drive in a population.
topic gene drive
maternal effect
HACK
mutagenic chain reaction
MCR
url http://g3journal.org/lookup/doi/10.1534/g3.116.034884
work_keys_str_mv AT chunchiehlin nonmendeliandominantmaternaleffectscausedbycrisprcas9transgeniccomponentsindrosophilamelanogaster
AT christopherjpotter nonmendeliandominantmaternaleffectscausedbycrisprcas9transgeniccomponentsindrosophilamelanogaster
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