Characterization of zebrafish Rad52 and replication protein A for oligonucleotide-mediated mutagenesis.

ABSTRACT: Zebrafish has become a favorite model organism not only in genetics and developmental biology, but also for the study of cancer, neuroscience and metabolism. However, strategies for reverse genetics in zebrafish are mostly limited to the use of antisense oligonucleotides, and therefore the development of other targeting methods is highly desirable. Here, we report an approach to gene targeting in this system in which single-stranded oligonucleotides and zebrafish Rad52 protein are employed. It has been proposed that a single-stranded oligonucleotide containing a mutation can be incorporated into the genome by annealing to the single-stranded region of the lagging strand of the replication fork. Rad52 is expected to accelerate the annealing step. In vitro experiments using purified truncate

SUBMITTER: Takahashi N 

PROVIDER: S-EPMC1182173 | biostudies-literature | 2005 Aug

REPOSITORIES: biostudies-literature