Failure to maintain full-term pregnancies in pig carrying klotho monoallelic knockout fetuses.

ABSTRACT:

Background

Small animals that show a deficiency in klotho exhibit extremely shortened life span with multiple aging-like phenotypes. However, limited information is available on the function of klotho in large animals such as pigs.

Results

In an attempt to produce klotho knockout pigs, an sgRNA specific for klotho (targeting exon 3) was designed and Cas9-sgRNA ribonucleoproteins were transfected into porcine fibroblasts. Transfected fibroblasts were cultured for one to 2 days and then directly used for nuclear transfer without selection. The cloned embryos were cultured in vitro for 7 days and analyzed to detect modifications of the klotho gene by both T7E1 and deep sequencing analysis. Modification succeeded in 13 of 20 blastocysts (65%), 8 of which (40.0%) were monoallelic

SUBMITTER: Lee S 

PROVIDER: S-EPMC7791653 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature