Genetic tools for the study of the mangrove killifish, Kryptolebias marmoratus, an emerging vertebrate model for phenotypic plasticity.

Autor: Li CY; Department of Biology, University of Maryland, College Park, Maryland, USA., Boldt H; Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.; Department of Orthopaedic Surgery, Boston Children's Hospital, Boston, Massachusetts, USA., Parent E; Department of Biology, University of Maryland, College Park, Maryland, USA., Ficklin J; Department of Biology, University of Maryland, College Park, Maryland, USA.; College of Computer, Mathematical, and Natural Sciences, Biological Sciences Graduate Program, University of Maryland, College Park, Maryland, USA., James A; Department of Orthopaedic Surgery, Boston Children's Hospital, Boston, Massachusetts, USA., Anlage TJ; Department of Biology, University of Maryland, College Park, Maryland, USA., Boyer LM; Department of Biology, University of Maryland, College Park, Maryland, USA., Pierce BR; Department of Biology, University of Maryland, College Park, Maryland, USA., Siegfried KR; Department of Biology, University of Massachusetts, Boston, Massachusetts, USA., Harris MP; Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.; Department of Orthopaedic Surgery, Boston Children's Hospital, Boston, Massachusetts, USA., Haag ES; Department of Biology, University of Maryland, College Park, Maryland, USA.
Jazyk: angličtina
Zdroj: Journal of experimental zoology. Part B, Molecular and developmental evolution [J Exp Zool B Mol Dev Evol] 2024 May; Vol. 342 (3), pp. 164-177. Date of Electronic Publication: 2023 Aug 08.
DOI: 10.1002/jez.b.23216
Abstrakt: Kryptolebias marmoratus (Kmar), a teleost fish of the order Cyprinodontiformes, has a suite of unique phenotypes and behaviors not observed in other fishes. Many of these phenotypes are discrete and highly plastic-varying over time within an individual, and in some cases reversible. Kmar and its interfertile sister species, K. hermaphroditus, are the only known self-fertile vertebrates. This unusual sexual mode has the potential to provide unique insights into the regulation of vertebrate sexual development, and also lends itself to genetics. Kmar is easily adapted to the lab and requires little maintenance. However, its internal fertilization and small clutch size limits its experimental use. To support Kmar as a genetic model, we compared alternative husbandry techniques to maximize recovery of early cleavage-stage embryos. We find that frequent egg collection enhances yield, and that protease treatment promotes the greatest hatching success. We completed a forward mutagenesis screen and recovered several mutant lines that serve as important tools for genetics in this model. Several will serve as useful viable recessive markers for marking crosses. Importantly, the mutant kissylips lays embryos at twice the rate of wild-type. Combining frequent egg collection with the kissylips mutant background allows for a substantial enhancement of early embryo yield. These improvements were sufficient to allow experimental analysis of early development and the successful mono- and bi-allelic targeted knockout of an endogenous tyrosinase gene with CRISPR/Cas9 nucleases. Collectively, these tools will facilitate modern developmental genetics in this fascinating fish, leading to future insights into the regulation of plasticity.
(© 2023 The Authors. Journal of Experimental Zoology Part B: Molecular and Developmental Evolution published by Wiley Periodicals LLC.)
Databáze: MEDLINE
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