Germline Editing of Drosophila Using CRISPR-Cas9-based Cytosine and Adenine Base Editors

0577609 - BC 2024 RIV US eng J - Článek v odborném periodiku
Thakkar, Nirav - Hejzlarová, Adéla - Brabec, Václav - Doležel, David
Germline Editing of Drosophila Using CRISPR-Cas9-based Cytosine and Adenine Base Editors.
CRISPR J. Roč. 6, č. 6 (2023), s. 557-569. ISSN 2573-1599. E-ISSN 2573-1602
Grant CEP: GA ČR(CZ) GA22-10088S
Institucionální podpora: RVO:60077344
Klíčová slova: Drosophila * base editors * Cas9 * germline editing * circadian clock * temperature
Obor OECD: Genetics and heredity (medical genetics to be 3)
Impakt faktor: 3.7, rok: 2022
Způsob publikování: Omezený přístup
https://www.liebertpub.com/doi/epub/10.1089/crispr.2023.0026

Target-AID, BE3, and ABE7.10 base editors fused to the catalytically modified Cas9 and xCas9(3.7) were tested for germline editing of the fruit fly Drosophila melanogaster. We developed a guide RNA-expressing construct, white-4gRNA, targeting splice sites in the white gene, an X-chromosome located gene. Using white-4gRNA flies and transgenic lines expressing Target-AID, BE3, and ABE7.10 base editors, we tested the efficiency of stable germline gene editing at three different temperatures. Classical Cas9 generating insertions/deletions by non-homologous end joining served as a reference. Our data indicate that gene editing is most efficient at 28 C, the highest temperature suitable for fruit flies. Finally, we created a new allele of the core circadian clock gene timeless using Target-AID. This base edited mutant allele timSS308-9FL had a disrupted circadian clock with a period of *29 h. The white-4gRNA expressing fly can be used to test new generations of base editors for future applications in Drosophila.
Trvalý link: https://hdl.handle.net/11104/0350058