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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60077344%3A_____%2F23%3A00577609" target="_blank" >RIV/60077344:_____/23:00577609 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/60076658:12310/23:43907454

Výsledek na webu

<a href="https://www.liebertpub.com/doi/epub/10.1089/crispr.2023.0026" target="_blank" >https://www.liebertpub.com/doi/epub/10.1089/crispr.2023.0026</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1089/crispr.2023.0026" target="_blank" >10.1089/crispr.2023.0026</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10603 - Genetics and heredity (medical genetics to be 3)

Projekt

<a href="/cs/project/GA22-10088S" target="_blank" >GA22-10088S: Funkční analýza proteinu Timeless</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2023

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

CRISPR J

ISSN

2573-1599

e-ISSN

2573-1602

Svazek periodika

6

Číslo periodika v rámci svazku

6

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

13

Strana od-do

557-569

Kód UT WoS článku

001130373700001

EID výsledku v databázi Scopus

2-s2.0-85176465060