A Comprehensive Toolbox for Genome Editing in Cultured Drosophila melanogaster Cells.

ABSTRACT: Custom genome editing has become an essential element of molecular biology. In particular, the generation of fusion constructs with epitope tags or fluorescent proteins at the genomic locus facilitates the analysis of protein expression, localization, and interaction partners at physiologic levels. Following up on our initial publication, we now describe a considerably simplified, more efficient, and readily scalable experimental workflow for PCR-based genome editing in cultured Drosophila melanogaster cells. Our analysis at the act5C locus suggests that PCR-based homology arms of 60 bp are sufficient to reach targeting efficiencies of up to 80% after selection; extension to 80 bp (PCR) or 500 bp (targeting vector) did not further improve the yield. We have expanded our targeting system to

SUBMITTER: Kunzelmann S 

PROVIDER: S-EPMC4889673 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature