Voutev Roumen, Mann Richard S
Departments of Biochemistry and Molecular Biophysics and Systems Biology, Columbia University, New York, NY.
Biotechniques. 2017 Jan 1;62(1):37-38. doi: 10.2144/000114494.
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 . To further diversify the available tools for genome engineering, we successfully harnessed the phage recombinase Bxb1 to perform recombinase-mediated cassette exchange (RMCE) in . 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.
快速且可靠的基因组修饰为任何基因组实体(基因、调控DNA、非编码RNA等)的详细体内功能分析提供了基础。随着CRISPR/Cas9基因组编辑技术的出现,对特定基因组位点的操作已成为包括果蝇在内的多种模式生物中的常规任务。为了进一步丰富基因组工程可用工具的种类,我们成功利用噬菌体重组酶Bxb1在……中进行重组酶介导的盒式交换(RMCE)。我们证明Bxb1具有高效的重组酶活性,可单独使用或与其他现有重组酶联合使用,以创建用于靶向位点盒式交换的平台。