Efficient gene editing of pig embryos by combining electroporation and lipofection.

Background And Aim: Mosaicism, which is characterized by the presence of wild-type and more than one mutant allele, poses a serious problem in zygotic gene modification through the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 system. Therefore, we used pig embryos to compare the gene editing efficiencies achieved by combining electroporation and lipofection using different aminopeptidase N (APN)-targeting guide RNA (gRNA) sequences.

Materials And Methods: Six gRNAs (gRNA1-6) with different target sequences were designed to target APN.

Impact of redox status of donor cows before superovulation treatment on in vivo embryo production.

This study investigated the relationship between oxidation and antioxidation parameters before superovulation (SOV) treatment and embryo recovery in donor cows. The relative redox status of the 61 donor cows was evaluated based on the median values of diacron-reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP) measurements (d-ROMs of 100 U.CARR; BAP of 2413  mol L ) before SOV treatment.

Effects of ergothioneine supplementation on meiotic competence and porcine oocyte development.

Background And Aim: The antioxidant effects of ergothioneine (EGT) on culture of porcine zygote are not well established. The study investigated the impact of EGT supplementation on meiotic competence and development of porcine oocytes.

Materials And Methods: The impact of EGT concentrations (0, 5, 10, 25, 50, and 100 μM) during maturation (IVM) on the progression of meiotic maturation, fertilization, and blastocyst formation in porcine oocytes was assessed.

Genome editing of porcine zygotes via lipofection of two guide RNAs using a CRISPR/Cas9 system.

CRISPR/Cas9-based multiplex genome editing via electroporation is relatively efficient; however, lipofection is versatile because of its ease of use and low cost. Here, we aimed to determine the efficiency of lipofection in CRISPR/Cas9-based multiplex genome editing using growth hormone receptor (GHR) and glycoprotein alpha-galactosyltransferase 1 (GGTA1)-targeting guide RNAs (gRNAs) in pig zygotes. Zona pellucida-free zygotes were collected 10 h after in vitro fertilization and incubated with Cas9, gRNAs, and Lipofectamine 2000 (LP2000) for 5 h.