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3. Demonstration ofin vivoengineered tandem duplications of varying sizes using CRISPR and recombinases inDrosophila melanogaster

Author

Loehlin, David W., McClain, Georgia L., Xu, Manting, Kedia, Ria, and Root, Elise

Subjects
Article
Abstract

Tandem gene duplicates are important parts of eukaryotic genome structure, yet the phenotypic effects of new tandem duplications are not well-understood, in part owing to a lack of techniques to build and modify them. We introduce a method, Recombinase-Mediated Tandem Duplication (RMTD), to engineer specific tandem duplicationsin vivousing CRISPR and recombinases. We describe construction of four different tandem duplications of theAlcohol Dehydrogenase(Adh) gene inDrosophila melanogaster, with duplicated block sizes ranging from 4.2 kb to 20.7 kb. Flies with theAdhduplications show elevated ADH enzyme activity over unduplicated single copies. This approach to engineering duplications is combinatoric, opening the door to systematic study of the relationship between the structure of tandem duplications and their effects on expression.

Published
2023

4. Demonstration of in vivo engineered tandem duplications of varying sizes using CRISPR and recombinases in Drosophila melanogaster.

Author

Loehlin DW, McClain GL, Xu M, Kedia R, and Root E

Subjects
Animals, Clustered Regularly Interspaced Short Palindromic Repeats, Genome, Drosophila melanogaster genetics, Gene Duplication
Abstract

Tandem gene duplicates are important parts of eukaryotic genome structure, yet the phenotypic effects of new tandem duplications are not well-understood, in part owing to a lack of techniques to build and modify them. We introduce a method, Recombinase-Mediated Tandem Duplication, to engineer specific tandem duplications in vivo using CRISPR and recombinases. We describe construction of four different tandem duplications of the Alcohol Dehydrogenase (Adh) gene in Drosophila melanogaster, with duplicated block sizes ranging from 4.2 to 20.7 kb. Flies with the Adh duplications show elevated ADH enzyme activity over unduplicated single copies. This approach to engineering duplications is combinatoric, opening the door to systematic study of the relationship between the structure of tandem duplications and their effects on expression., Competing Interests: Conflicts of interest statement The authors declare no conflict of interest., (© The Author(s) 2023. Published by Oxford University Press on behalf of The Genetics Society of America.)

Published
2023
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