High-efficiency Non-mosaic CRISPR-mediated Knock-in and Indel Mutation in F0

Overview

Journal Development

Specialty Biology

Date 2017 Jul 12

PMID 28694259

Citations 34

Authors

Yetki Aslan

Emmanuel Tadjuidje

Aaron M Zorn

Sang-Wook Cha

Affiliations

Soon will be listed here.

Abstract

The revolution in CRISPR-mediated genome editing has enabled the mutation and insertion of virtually any DNA sequence, particularly in cell culture where selection can be used to recover relatively rare homologous recombination events. The efficient use of this technology in animal models still presents a number of challenges, including the time to establish mutant lines, mosaic gene editing in founder animals, and low homologous recombination rates. Here we report a method for CRISPR-mediated genome editing in oocytes with homology-directed repair (HDR) that provides efficient non-mosaic targeted insertion of small DNA fragments (40-50 nucleotides) in 4.4-25.7% of F0 tadpoles, with germline transmission. For both CRISPR/Cas9-mediated HDR gene editing and indel mutation, the gene-edited F0 embryos are uniformly heterozygous, consistent with a mutation in only the maternal genome. In addition to efficient tagging of proteins , this HDR methodology will allow researchers to create patient-specific mutations for human disease modeling in .

Citing Articles

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