biostudies-literatureBansal ADepartment of Biotechnology, Ministry of Science and Technology, India24076https://www.ebi.ac.uk/biostudies/studies/S-EPMC11473785biostudies-literatureUnknown14(1)Conditional regulation is a highly beneficial system for studying the function of essential genes in Plasmodium falciparum and dimerizable Cre recombinase (DiCre) is a recently adapted conditional regulation system suitable for this purpose. In the DiCre system, two inactive fragments of Cre are reconstituted to form a functionally active enzyme in the presence of rapamycin. Different loci have been targeted to generate parasite lines that express the DiCre enzyme. Here, we have used marker-free CRISPR-Cas9 gene editing to integrate the DiCre cassette in a redundant cg6 locus. We have shown the utility of the newly generated ∆cg6DC4 parasites in mediating robust, rapid, and highly specific excision of exogenously encoded gfp sequence. The ∆cg6DC4 parasites are also capable of conditional eScientific reportsGeneration of a new DiCre expressing parasite strain for functional characterization of Plasmodium falciparum genes in blood stages.PMC11473785BT/PR38411/GET/119/311/2020Bansal AChoudhury HSharma MfalseGeneration of a new DiCre expressing parasite strain for functional characterization of Plasmodium falciparum genes in blood stages.Conditional regulation is a highly beneficial system for studying the function of essential genes in Plasmodium falciparum and dimerizable Cre recombinase (DiCre) is a recently adapted conditional regulation system suitable for this purpose. In the DiCre system, two inactive fragments of Cre are reconstituted to form a functionally active enzyme in the presence of rapamycin. Different loci have been targeted to generate parasite lines that express the DiCre enzyme. Here, we have used marker-free CRISPR-Cas9 gene editing to integrate the DiCre cassette in a redundant cg6 locus. We have shown the utility of the newly generated ∆cg6DC4 parasites in mediating robust, rapid, and highly specific excision of exogenously encoded gfp sequence. The ∆cg6DC4 parasites are also capable of conditional e2024-01-01T00:00:00Z2024 Oct2025-04-04T02:19:56.149Z2025-04-04T02:19:56.149ZS-EPMC114737853940238010.1038/s41598-024-75657-x