Source: UNIVERSITY OF CALIFORNIA, DAVIS submitted to
GERM CELL AND EMBRYO DEVELOPMENT AND MANIPULATION FOR THE IMPROVEMENT OF LIVESTOCK
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1021308
Grant No.
(N/A)
Project No.
CA-D-ASC-2070-RR
Proposal No.
(N/A)
Multistate No.
W-4171
Program Code
(N/A)
Project Start Date
Dec 24, 2019
Project End Date
Oct 13, 2020
Grant Year
(N/A)
Project Director
Ross, PA, JU.
Recipient Organization
UNIVERSITY OF CALIFORNIA, DAVIS
410 MRAK HALL
DAVIS,CA 95616-8671
Performing Department
Animal Science
Non Technical Summary
This project supports the mission of the Agricultural Experiment Station by addressing the Hatch Act area(s) of: plant and animal production, protection, and health; molecular biology; biotechnology.Generation of genetically enhanced animals for the production of food and fiber holds significant promise for consumers, animal producers, their communities and our environment. Conventional and somatic cell nuclear transfer (SCNT), or cloning, has dramatically advanced animal biotechnology, significantly enhancing our ability to produce genetically altered livestock. However the current technology is very inefficient therefore limiting its broad application. The cause of this inefficiency is broadly assigned to incomplete nuclear reprogramming of the donor cell upon nuclear transplantation. The mechanisms of nuclear reprogramming are largely uncharacterized. Studies will be conducted to determine the mechanisms of epigenetic reprogramming during normal embryonic development and that knowledge will be applied to facilitate nuclear reprogramming after somatic cell nuclear transfer with the objective of improving the efficiency of this technology.
Animal Health Component
0%
Research Effort Categories
Basic
80%
Applied
20%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3013910104050%
3013910105050%
Knowledge Area
301 - Reproductive Performance of Animals;

Subject Of Investigation
3910 - Cross-commodity research--multiple animal species;

Field Of Science
1050 - Developmental biology; 1040 - Molecular biology;
Goals / Objectives
Understand the biology of gamete development, fertilization and embryogenesis including the underlying cellular and molecular mechanisms.
Project Methods
My laboratory will contribute to objective 1 by conducting research oriented to elucidate the mechanism of epigenetic reprogramming that the paternal genomes undergo after fertilization to form the totipotent zygote, and how this epigenetic reprogramming leads to activation of the embryonic genome.

Progress 12/24/19 to 10/13/20

Outputs
Target Audience:Undergraduate, graduate, and post-doctoral students, livestock producers, general public and the scientific community. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project provided graduate student training for 4 PhD students and two postdoctoral fellow. Furthermore, several undergraduate students participated in the research. How have the results been disseminated to communities of interest?Results were presented at meetings which gather embryo transfer practitioners, companies, and scientists interested in embryos and gene editing technologies. Talks were delivered to students, colleagues and practitioners at the International Embryo Technologies Society. Papers were published in scientific journals with widespread dissemination. What do you plan to do during the next reporting period to accomplish the goals?This is the final report

Impacts
What was accomplished under these goals? Relevant to the objective of understanding the biology and underlying mechanisms of gamete development, fertilization, and embryogenesis, we have: 1) Identified changes in chromatin accessibility across the bovine genome during preimplantation embryo development. Bioinformatics analysis of this data reveled potential regulators of chromatin remodeling during bovine preimplantation development. 2) Identified transcripts activated during bovine major EGA, as well as potential regulators involved in activation of the genome and degradation of maternal transcripts. 3) Derived and cultured sheep embryonic stem cells. 4) Developed and effective electroporation protocol for CRISPR/Cas9 based gene editing of cattle zygotes. 4) We reviewed the literature related to IVF and embryo cryopreservation.

Publications

  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Halstead MM, Ma X, Schultz RM, Ross PJ. Chromatin remodeling in bovine embryos indicates species-specific regulation of genome activation. Nature Communications. 2020; 11:4654
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Bogliotti YS, Chung N, Paulson E, Chitwood J, Halstead M, Kern C, Schultz RM, Ross PJ. Transcript profiling of bovine embryos implicates specific transcription factors in the maternal-to-embryo transition. Biol Reprod 2020 Mar 13;102(3):671-679. doi: 10.1093/biolre/ioz209.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Vilarino M, Soto DA, Bogliotti YS, Yu L, Zhang Y, Wang C, Paulson E, Zhong C. Jin M, Izpisua Belmonte JC, Wu J, Ross PJ. Derivation of sheep embryonic stem cells under optimized conditions. Reproduction 2020; 160(5):761-772. https://doi.org/10.1530/REP-19-0606
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Camargo LSA, Owen JR, Van Eenennaam AL, Ross PJ. Efficient One-Step Knockout by Electroporation of Ribonucleoproteins into Zona-Intact Bovine Embryos. Frontiers in Genetics 2020 | https://doi.org/10.3389/fgene.2020.570069
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Ferr� LB, Kjelland ME, Taiyeb AM, Campos-Chillon LF, Ross PJ. Recent Progress in Bovine In Vitro-Derived Embryo Cryotolerance: Impact of In Vitro Culture Systems, Advances in Cryopreservation and Future Considerations. Reprod Domest Anim. 2020 Mar 7. doi: 10.1111/rda.13667.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Navarro M, Soto DA, Pinzon CA, Wu J, Ross PJ. Livestock pluripotency is finally captured in vitro. Reprod Fert Dev 2020; 32(2) 11-39 https://doi.org/10.1071/RD19272
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Ferr� LB, Kjelland ME, Str�bech LB, Hyttel P, Mermillod P, Ross PJ. Recent advances in bovine in vitro embryo production: reproductive biotechnology history and methods. Animal. 2020 May;14(5):991-1004. doi: 10.1017/S1751731119002775.