Class

Article

College

College of Agriculture and Applied Sciences

Department

Animal, Dairy, and Veterinary Sciences Department

Presentation Type

Oral Presentation

Abstract

While a useful artificial reproduction technology, the method of somatic cell nuclear transfer (scNT) is generally inefficient, with successful pregnancy rates typically below 10%. Following nuclear transfer, the cellular machinery of the oocyte must reprogram the somatic DNA for proper embryo development. The maternal to zygotic transition (MZT) is associated with marked changes in control of gene expression, as well as marked class changes in small non-coding RNAs (sncRNAs), as previously identified by our research group in bovine embryos. Failure for these class changes to occur in scNT embryos may drive higher failure rates. The present study is the first to employ a discovery-based, small RNA sequencing approach to determine the population of sncRNAs in bovine scNT embryos as compared to in vitro-fertilized embryos. In all populations of sncRNA surveyed, large-scale population differences between ScNT and IVF embryos were not apparent for miRNAs, piRNAs, or tRNA fragments. However, we did identify a few select miRNAs that were differentially expressed at specific stages of embryo development, including miR-2340-3p and miR-345-5p in the morula-staged embryos, as well as miR-497-5p at the blastocyst-staged embryos. Distinct populations of piRNA-like RNAs (pilRNAs) were identified through development of the scNT embryos, and abundance of several species were differentially expressed in scNT versus IVF embryos at the morula and blastocyst stages. Transfer RNA fragments may function in RNA interference pathways similar to miRNA, and we identified 6 tRNAs differentially expressed in scNT and IVF embryos during early embryo development. In conclusion, widespread sncRNA population differences were not observed between IVF and ScNT embryos, suggesting that errors associated with scNT may be somewhat random in nature. However, those sncRNAs that were consistently differently expressed in scNT embryos may be important contributors to successful genome reprogramming of cloned embryos and these targets warrant further functional analyses.

Location

Room 155

Start Date

4-10-2019 12:00 PM

End Date

4-10-2019 1:15 PM

Included in

Life Sciences Commons

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Apr 10th, 12:00 PM Apr 10th, 1:15 PM

Somatic Cell Nuclear Transfer in Early Bovine Embryo Development is Associated with Changes in Small Non-Coding RNA Species

Room 155

While a useful artificial reproduction technology, the method of somatic cell nuclear transfer (scNT) is generally inefficient, with successful pregnancy rates typically below 10%. Following nuclear transfer, the cellular machinery of the oocyte must reprogram the somatic DNA for proper embryo development. The maternal to zygotic transition (MZT) is associated with marked changes in control of gene expression, as well as marked class changes in small non-coding RNAs (sncRNAs), as previously identified by our research group in bovine embryos. Failure for these class changes to occur in scNT embryos may drive higher failure rates. The present study is the first to employ a discovery-based, small RNA sequencing approach to determine the population of sncRNAs in bovine scNT embryos as compared to in vitro-fertilized embryos. In all populations of sncRNA surveyed, large-scale population differences between ScNT and IVF embryos were not apparent for miRNAs, piRNAs, or tRNA fragments. However, we did identify a few select miRNAs that were differentially expressed at specific stages of embryo development, including miR-2340-3p and miR-345-5p in the morula-staged embryos, as well as miR-497-5p at the blastocyst-staged embryos. Distinct populations of piRNA-like RNAs (pilRNAs) were identified through development of the scNT embryos, and abundance of several species were differentially expressed in scNT versus IVF embryos at the morula and blastocyst stages. Transfer RNA fragments may function in RNA interference pathways similar to miRNA, and we identified 6 tRNAs differentially expressed in scNT and IVF embryos during early embryo development. In conclusion, widespread sncRNA population differences were not observed between IVF and ScNT embryos, suggesting that errors associated with scNT may be somewhat random in nature. However, those sncRNAs that were consistently differently expressed in scNT embryos may be important contributors to successful genome reprogramming of cloned embryos and these targets warrant further functional analyses.