Replacing sperm with genotyped haploid androgenetic blastomeres to generate cattle with predetermined paternal genomes

• Published in: Biology of reproduction 2024-09, Vol.111 (6), p.1311
• Main Authors: Aguila, Luis, Sampaio, Rafael V, Therrien, Jacinthe, Nociti, Ricardo P, Labrecque, Remi, Tremblay, Amelie, Marras, Gabrielle, Blondin, Patrick, Smith, Lawrence C
• Format: Article
• Language: English
• Subjects: Animals; Blastomeres - cytology; Blastomeres - metabolism; Cattle - genetics; Cloning, Organism - methods; Cloning, Organism - veterinary; Embryonic Development - genetics; Female; Genome; Genotype; Haploidy; Male; Pregnancy; Sperm Injections, Intracytoplasmic; Spermatozoa
• ISSN: 0006-3363; 1529-7268; 1529-7268
• DOI: 10.1093/biolre/ioae135

Although meiosis plays an essential role for the survival of species in natural selection, the genetic diversity resulting from sexual reproduction impedes human-driven strategies to transmit the most suitable genomes for genetic improvement, forcing breeders to select diploid genomes generated after fertilization, that is, after the encounter of sperm and oocytes carrying unknown genomes. To determine whether genomic assessment could be used before fertilization, some androgenetic haploid morula-stage bovine embryos derived from individual sperm were biopsied for genomic evaluation and others used to reconstruct "semi-cloned" (SC) diploid zygotes by the intracytoplasmic injection into parthenogenetically activated oocytes, and the resulting embryos were transferred to surrogate females to obtain gestations. Compared to controls, in vitro development to the blastocyst stage was lower and fewer surrogates became pregnant from the transfer of SC embryos. However, fetometric measurements of organs and placental membranes of all SC conceptuses were similar to controls, suggesting a normal post-implantation development. Moreover, transcript amounts of imprinted genes IGF2, IGF2R, PHLDA2, SNRPN, and KCNQ1OT1 and methylation pattern of the KCNQ1 DMR were unaltered in SC conceptuses. Overall, this study shows that sperm can be replaced by genotyped haploid embryonic-derived cells to produce bovine embryos carrying a predetermined paternal genome and viable first trimester fetuses after transfer to female recipients. Haploid morula-stage embryonic cells derived from a single sperm can be genotyped and injected into activated oocytes to reconstruct diploid zygotes that develop both in vitro into blastocysts and in vivo into viable post-implantation bovine conceptuses with predetermined paternal genomes.