Optimization of a DiCre recombinase system with reduced leakage for conditional genome editing of Cryptosporidium.

ABSTRACT:

Background

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.

Methods

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 ch

SUBMITTER: Huang Y 

PROVIDER: S-EPMC11337648 | biostudies-literature | 2024 Aug

REPOSITORIES: biostudies-literature