A DREaMR system to simplify combining mutations with rescue transgenes in Aedes aegypti

TitleA DREaMR system to simplify combining mutations with rescue transgenes in Aedes aegypti
Publication TypeJournal Article
Year of Publication2021
AuthorsChen J, Luo J, Gurav AS, Chen Z, Wang Y, Montell C
JournalGenetics
Volume219
Issue3
Date Published2021 11 05
ISSN1943-2631
KeywordsAedes, Animals, Animals, Genetically Modified, CRISPR-Cas Systems, Drosophila melanogaster, Gene Editing, Genes, Insect, Mosquito Control, Mosquito Vectors, Mutation
Abstract

In most experimental animals, it is challenging to combine mutations and rescue transgenes and to use bipartite systems to assess gene expression. To circumvent the difficulties in combining multiple genetic elements, we developed the DREaMR (Drug-on, REporter, Mutant, Rescue) system. Using Drosophila white as the initial model, we demonstrated that introduction of a single insertion by CRISPR/Cas9 created a null mutation, a tagged rescue construct, which could be induced with doxycycline, and which allowed assessment of protein expression. To create a DREaMR in an organism in which combining multiple genetic elements is more problematic than in Drosophila, we tested the mosquito, Aedes aegypti-the insect vector for dengue, yellow fever, Zika, and other viral diseases. We generated a DREaMR allele in the kh gene, which permitted us to induce expression of the rescue construct, and detect expression of Kh. Thus, this system avoids the need to perform genetic crosses to introduce an inducible rescue transgene in a mutant background, or to combine driver and reporter lines to examine expression of the targeted protein. We propose that DREaMR provides a system that can be applied to additional mosquito vectors as well as other organisms in which CRISPR/Cas9 is effective.

DOI10.1093/genetics/iyab146
Alternate JournalGenetics
PubMed ID34740249
PubMed Central IDPMC8570792
Grant ListR01 AI165575 / AI / NIAID NIH HHS / United States
R01 DC007864 / DC / NIDCD NIH HHS / United States
R01 EY008117 / EY / NEI NIH HHS / United States
R01 EY010852 / EY / NEI NIH HHS / United States