Intronic gRNAs for the construction of minimal gene drive systems
File(s)fbioe-10-857460.pdf (1.54 MB)
Published version
Author(s)
Nash, Alexander
Paolo, Capriotti
Hoermann, Astrid
Papathanos, Phillipos
Windbichler, Nikolai
Type
Journal Article
Abstract
Gene drives are promising tools for the genetic control of insect vector or pest populations. CRISPR-based gene drives are generally highly complex synthetic constructs consisting of multiple transgenes and their respective regulatory elements. This complicates the generation of new gene drives and the testing of the behavior of their constituent functional modules. Here, we explored the minimal genetic components needed to constitute autonomous gene drives in Drosophila melanogaster. We first designed intronic gRNAs that can be located directly within coding transgene sequences and tested their functions in cell lines. We then integrated a Cas9 open reading frame hosting such an intronic gRNA within the Drosophila rcd-1r locus that drives the expression in the male and female germlines. We showed that upon removal of the fluorescent transformation marker, the rcd-1rd allele supports efficient gene drive. We assessed the propensity of this driver, designed to be neutral with regards to fitness and host gene function, to propagate in caged fly populations. Because of their simplicity, such integral gene drives could enable the modularization of drive and effector functions. We also discussed the possible biosafety implications of minimal and possibly recoded gene drives.
Date Issued
2022-05-12
Date Acceptance
2022-03-24
Citation
Frontiers in Bioengineering and Biotechnology, 2022, 10, pp.1-10
ISSN
2296-4185
Publisher
Frontiers Media
Start Page
1
End Page
10
Journal / Book Title
Frontiers in Bioengineering and Biotechnology
Volume
10
Copyright Statement
© 2022 Nash, Capriotti, Hoermann, Papathanos and Windbichler. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
License URL
Identifier
https://www.frontiersin.org/articles/10.3389/fbioe.2022.857460/full
Subjects
Drosophila
gene drives
genetic control
genetics
synthetic biology
0699 Other Biological Sciences
0903 Biomedical Engineering
1004 Medical Biotechnology
Publication Status
Published
Article Number
857460
Date Publish Online
2022-05-12