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Improved Growth and High Inheritance of Melanocortin-4 Receptor (mc4r) Mutation in CRISPR/Cas-9 Gene-Edited Channel Catfish, Ictalurus punctatus

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Abstract

Effects of CRISPR/Cas9 knockout of the melanocortin-4 receptor (mc4r) gene in channel catfish, Ictalurus punctatus, were investigated. Three sgRNAs targeting the channel catfish mc4r gene in conjunction with Cas9 protein were microinjected in embryos and mutation rate, inheritance, and growth were studied. Efficient mutagenesis was achieved as demonstrated by PCR, Surveyor® assay, and DNA sequencing. An overall mutation rate of 33% and 33% homozygosity/bi-allelism was achieved in 2017. Approximately 71% of progeny inherited the mutation. Growth was generally higher in MC4R mutants than controls (CNTRL) at all life stages and in both pond and tank environments. There was a positive relationship between zygosity and growth, with F1 homozygous/bi-allelic mutants reaching market size 30% faster than F1 heterozygotes in earthen ponds (p = 0.022). At the stocker stage (~ 50 g), MC4R × MC4R mutants generated in 2019 were 40% larger than the mean of combined CNTRL × CNTRL families (p = 0.005) and 54% larger than F1 MC4R × CNTRL mutants (p = 0.001) indicating mutation may be recessive. With a high mutation rate and inheritance of the mutation as well as improved growth, the use of gene-edited MC4R channel catfish appears to be beneficial for application on commercial farms.

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Funding

This work was funded by the United States Department of Agriculture National Institute of Food and Agriculture award 2015–67015-23488 to Roger Cone and sub-award to Rex Dunham.

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Authors and Affiliations

  1. Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL, 36849, USA

    Michael Coogan, Veronica Alston, Baofeng Su, Karim Khalil, Ahmed Elaswad, Mohd Khan, Andrew Johnson, De Xing, Shangjia Li, Jinhai Wang, Rhoda M. C. Simora, Cuiyu Lu, Wenwen Wang, Darshika Hettiarachchi, Tasnuba Hasin, Ian A. E. Butts & Rex A. Dunham

  2. Department of Animal Wealth Development, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, 41522, Egypt

    Ahmed Elaswad

  3. Department of Fisheries Biology and Genetics, Agricultural University, Mymensingh, 2202, Bangladesh

    Mohd Khan

  4. College of Fisheries and Ocean Sciences, University of the Philippines Visayas, 5023, Miagao, Iloilo, Philippines

    Rhoda M. C. Simora

  5. Department of Molecular and Integrative Physiology, Vanderbilt University, Nashville, TN, 37203-5721, USA

    Patrick Page-McCaw, Wenbiao Chen, Max Michel & Roger D. Cone

Authors
  1. Michael Coogan
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  2. Veronica Alston
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  21. Rex A. Dunham
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Contributions

Conceptualization: Michael Coogan, Rex Dunham, Karim Khalil, Ahmed Elaswad, Mohd Khan, Roger D. Cone; methodology: Michael Coogan, Rex Dunham, Karim Kalil, Ahmed Elaswad, Patrick Page-McCaw, Wenbiao Chen, Maximilian Michel, Mohd Khan; formal analysis and investigation: Michael Coogan; writing—original draft preparation: Michael Coogan; writing—review and editing: Michael Coogan, Veronica Alston, Baofeng Su, Rex Dunham, Ian Butts, Andrew Johnson, De Xing, Jinhai Wang, Shangjia Li, Rhoda M.C. Simora, Wenwen Wang, Darshika Hettiarachchi, Cuiyu Lu; funding acquisition: Rex Dunham, Roger D. Cone; Resources: Rex Dunham; supervision: Rex Dunham, Ian Butts; data curation: Andrew Johnson, De Xing, Jinhai Wang, Shangjia Li, Rhoda M.C. Simora, Wenwen Wang, Darshika Hettiarachchi, Cuiyu Lu, and Tasnuba Hasin.

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Correspondence to Michael Coogan.

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Coogan, M., Alston, V., Su, B. et al. Improved Growth and High Inheritance of Melanocortin-4 Receptor (mc4r) Mutation in CRISPR/Cas-9 Gene-Edited Channel Catfish, Ictalurus punctatus. Mar Biotechnol 24, 843–855 (2022). https://doi.org/10.1007/s10126-022-10146-8

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