biostudies-literatureHuang YDouble First-Class Discipline Promotion Project111 ProjectNational Natural Science Foundation of ChinaGuangdong Major Project of Basic and Applied Basic ResearchNational Key Research and Development Program of China352https://www.ebi.ac.uk/biostudies/studies/S-EPMC11337648biostudies-literatureUnknown17(1)<h4>Background</h4>The dimerizable Cre recombinase system (DiCre) exhibits increased leaky activity in Cryptosporidium, leading to unintended gene editing in the absence of induction. Therefore, optimization of the current DiCre technique is necessary for functional studies of essential Cryptosporidium genes.<h4>Methods</h4>Based on the results of transcriptomic analysis of Cryptosporidium parvum stages, seven promoters with different transcriptional capabilities were screened to drive the expression of Cre fragments (FKBP-Cre59 and FRB-Cre60). Transient transfection was performed to assess the effect of promoter strength on leakage activity. In vitro and in vivo experiments were performed to evaluate the leaky activity and cleavage efficiency of the optimized DiCre system by polymerase chParasites & vectorsOptimization of a DiCre recombinase system with reduced leakage for conditional genome editing of Cryptosporidium.PMC11337648D200082022YFD18002002023B10564003320301092020B0301030007Liu HPei SLi JHuang YXiao LGuo YXu RFeng YYou HLi NfalseOptimization of a DiCre recombinase system with reduced leakage for conditional genome editing of Cryptosporidium.<h4>Background</h4>The dimerizable Cre recombinase system (DiCre) exhibits increased leaky activity in Cryptosporidium, leading to unintended gene editing in the absence of induction. Therefore, optimization of the current DiCre technique is necessary for functional studies of essential Cryptosporidium genes.<h4>Methods</h4>Based on the results of transcriptomic analysis of Cryptosporidium parvum stages, seven promoters with different transcriptional capabilities were screened to drive the expression of Cre fragments (FKBP-Cre59 and FRB-Cre60). Transient transfection was performed to assess the effect of promoter strength on leakage activity. In vitro and in vivo experiments were performed to evaluate the leaky activity and cleavage efficiency of the optimized DiCre system by polymerase ch2024-01-01T00:00:00Z2024 Aug2026-06-02T18:36:37.45Z2024-11-21T10:58:11.915ZS-EPMC113376483916943010.1186/s13071-024-06431-1