Multiple gene editing in porcine embryos using a combination of microinjection, electroporation, and transfection methods.

ABSTRACT:

Background and aim

Mosaicism - the presence of both wild-type and mutant alleles - is a serious problem for zygotic gene modification through gene editing using the Clustered regularly interspaced short palindromic repeats-Cas9 (CRISPR/Cas9) system. Different delivery methods, such as microinjection (MI), electroporation (EP), and transfection (TF), can be used to transfer CRISPR/Cas9 components into porcine zygotes. This study aimed to develop a method that combines MI, EP, and TF to improve mutation efficiency mediated through the CRISPR/Cas9 system for a triple-gene knockout in pigs.

Materials and methods

The study consisted of three groups: The MI group with three simultaneously microinjected guide RNAs (gRNAs) targeting α-1,3-galactosyltransferase (GGTA1), cytidi

SUBMITTER: Le QA 

PROVIDER: S-EPMC9631378 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature