Efficient genome editing using CRISPR/Cas9 ribonucleoprotein approach in cultured Medaka fish cells.

ABSTRACT: Gene editing with CRISPR/Cas9 is a powerful tool to study the function of target genes. Although this technology has demonstrated wide efficiency in many species, including fertilized zebrafish and medaka fish embryos when microinjected, its application to achieve efficient gene editing in cultured fish cells have met some difficulty. Here, we report an efficient and reliable approach to edit genes in cultured medaka (Oryzias latipes) fish cells using pre-formed gRNA-Cas9 ribonucleoprotein (RNP) complex. Both medaka fish haploid and diploid cells were transfected with the RNP complex by electroporation. Efficient gene editing was demonstrated by polymerase chain reaction (PCR) amplification of the target gene from genomic DNA and heteroduplex mobility assay carried out with polyacry

SUBMITTER: Liu Q 

PROVIDER: S-EPMC6124564 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature