BackgroundC31 integrase, a site-specific recombinase, can efficiently target attB-bearing transgenes to endogenous pseudo attP sites within mammalian genomes. The sequence features of endogenous binding sites will help us to fully understand the site-specific recognition function by C31 integrase. The present study was aimed to uncover the global map of phi C31 integrase binding sites in bovine cells and analysis the features of these binding sites by comprehensive bioinformatics methods.Results In this study, we constructed a ChIP-seq method that can be used to uncover the global binding sites by phiC31 integrase. 6740 potential phi C31 integrase binding sites were identified. A sequence motif was found that contains inverted repeats and has similarities to wild-type attP site. Using REPEATMASKER, we identified a total of 20,183 repeat-regions distributed in 50 repeat types for the 6740 binding sites. These sites enriched in regulation of GTPase activity of in the GO category of biological process and KEGG pathway of signal transmembrane transporter activity.ConclusionThis study is the first time to uncover the global map of binding sites for phi C31 integrase using ChIP-sequencing method and analysis the features of these binding sites. This method will help us to fully understand the mechanism of the site-specific integration function by phiC31 integrase and will potentially boost its genetic manipulations in both gene therapy and generation of transgenic animals.
[1]Shanghai Eighth Peoples Hosp, Dept Lab Med, Shanghai 200040, Peoples R China;
[2]Shanghai Jiao Tong Univ, Shanghai First Peoples Hosp, Dept Resp Med, Sch Med, Shanghai 201620, Peoples R China;
[3]Fudan Univ, Shanghai Inst Planned Parenthood Res, Lab Reprod Pharmacol, NHC Key Lab Reprod Regulat, Shanghai 200032, Peoples R China
(8.0+极速模式)