Bxb1 phage recombinase assists genome engineering in Drosophila melanogaster.
نویسندگان
چکیده
Rapid and reliable genome modifications provide the basis for detailed in vivo functional analysis of any genomic entity (gene, regulatory DNA, non-coding RNA, etc). With the advent of CRISPR/Cas9 genome editing technology, manipulation of a particular genomic locus has become a routine undertaking in variety of model organisms, including the fruit fly Drosophila melanogaster. To further diversify the available tools for genome engineering, we successfully harnessed the phage recombinase Bxb1 to perform recombinase-mediated cassette exchange (RMCE) in D. melanogaster. We demonstrate that Bxb1 possesses highly efficient recombinase activity and could be used alone or in conjunction with other currently available recombinases for creating platforms for cassette exchange of targeted loci.
منابع مشابه
Mycobacteriophage Bxb1 integrates into the Mycobacterium smegmatis groEL1 gene.
Mycobacteriophage Bxb1 is a temperate phage of Mycobacterium smegmatis and forms stable lysogens in which the Bxb1 genome is integrated into the host chromosome. Bxb1 encodes an integrase of the large serine recombinase family that catalyses integration and excision of the Bxb1 genome. We show here that Bxb1 integrates into a chromosomal attB site located within the 3' end of the groEL1 gene su...
متن کاملAccuracy and efficiency define Bxb1 integrase as the best of fifteen candidate serine recombinases for the integration of DNA into the human genome
BACKGROUND Phage-encoded serine integrases, such as φC31 integrase, are widely used for genome engineering. Fifteen such integrases have been described but their utility for genome engineering has not been compared in uniform assays. RESULTS We have compared fifteen serine integrases for their utility for DNA manipulations in mammalian cells after first demonstrating that all were functional ...
متن کاملThe Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis
BACKGROUND The mycobacteriophage large serine recombinase Bxb1 catalyzes site-specific recombination between its corresponding attP and attB recognition sites. Previously, we and others have shown that Bxb1 has catalytic activity in various eukaryotic species including Nicotiana tabacum, Schizosaccharomyces pombe, insects and mammalian cells. RESULTS In this work, the Bxb1 recombinase gene wa...
متن کاملControl of Phage Bxb1 Excision by a Novel Recombination Directionality Factor
Mycobacteriophage Bxb1 integrates its DNA at the attB site of the Mycobacterium smegmatis genome using the viral attP site and a phage-encoded integrase generating the recombinant junctions attL and attR. The Bxb1 integrase is a member of the serine recombinase family of site-specific recombination proteins and utilizes small (<50 base pair) substrates for recombination, promoting strand exchan...
متن کاملCaptured segment exchange: a strategy for custom engineering large genomic regions in Drosophila melanogaster.
Site-specific recombinases (SSRs) are valuable tools for manipulating genomes. In Drosophila, thousands of transgenic insertions carrying SSR recognition sites have been distributed throughout the genome by several large-scale projects. Here we describe a method with the potential to use these insertions to make custom alterations to the Drosophila genome in vivo. Specifically, by employing rec...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- BioTechniques
دوره 62 1 شماره
صفحات -
تاریخ انتشار 2017