To date, only porcine and bovine zygotes have been successfully electroporated to produce knockout live animals. Pig researchers have electroporated zygotes and oocytes to generate genome edited blastocysts and live piglets using Cas9 genome editing reagents.

Co-injection of zygotes with Cas9 mRNA and sgRNA has been proven to be an efficient gene-editing strategy for genome modification of different species. Genetic engineering in pigs holds a great promise in biomedical research.

Jan 31, 2014 · Here we report the efficient generation of biallelic knockout pigs in one step by direct cytoplasmic injection of Cas9 mRNA and sgRNA into zygotes.

Oct 10, 2013 · In this study we demonstrate that both TALEN and ZFN technology can be efficiently applied to engineer pig zygotes that result in gene edited live births, both mono- and bi-allelic (Table 1;...

Oct 9, 2016 · Here, we successfully achieved precise DMD targeting in Chinese Diannan miniature pigs by co-injecting zygotes with Cas9 mRNA and sgRNA targeting DMD. Two piglets were obtained after embryo transfer, one of piglets was identified as DMD -modified individual via traditional cloning, sequencing and T7EN1 cleavage assay.

Oct 10, 2013 · Transcription activator-like effector nuclease (TALEN) and zinc finger nuclease (ZFN) genome editing technology enables site directed engineering of the genome. Here we demonstrate for the first time that both TALEN and ZFN injected directly into pig zygotes can produce live genome edited pigs.

The increasing focus on the pig as a biomedical model calls for studies which investigate morphological and molecular mechanisms during initial embryonic development in this species. In the pig, the paternal genome is actively demethylated in the zygote, whereas the maternal genome remains methylate …

May 4, 2017 · Here, we review methods to optimize the developmental competence of genome-edited porcine embryos and strategies to improve the zygote genome-editing efficiency in pigs.

Jan 6, 2021 · Cytoplasmic microinjection and electroporation of the CRISPR/Cas9 system into zygotes are used for generating genetically modified pigs. However, these methods create mosaic mutations in embryos.

1 day ago · Schematic representation of generation of 8-gene-edited cloned pigs and their verification by pig to non-human primate kidney xenotransplantation.