Genome-wide genetic mutations accumulated in genome-edited pigs and their filial generation for xenotransplantation

Abstract

Although xenotransplantation has revolutionarily developed based on the usage of genome-edited pigs, it is still unknown whether these pigs and their offspring remains genomic stable. Here, we show that GGTA1-knockout (GTKO) pigs accumulated an average of 1205 genome-wide genetic mutations, and their filial 1 offspring contained an average of 18 de novo mutations compared with wild control and their parents, respectively. The majority of mutations were annotated in intergenic regions without altering protein functions and predicted off-target mutations. RNA-sequencing analysis and phenotype observation indicated that accumulated mutations may have a limited influence on GTKO pigs and most genetic mutations in GTKO pigs can be explained by electrotransfection of plasmids into cells. This is the first report that the genetic mutations of genome-edited pigs are inherited stably by their filial generation and provide a reference for safe application and the standard approach to breeding genome-edited pigs for xenotransplantation.