Identifying Conserved CRISPR Leader Motifs that regulate CRISPR Adaptive Immunity to RNA
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Bioinformatics
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This project investigates Cas6/RT/Cas1 protein fusions across various bacterial genomes, analyzing their CRISPR loci and leader sequences. Using bioinformatics tools including BLAST, CRISPRDetectv3, CRISPRLeaderExtractor, and MEME, we identified conserved DNA motifs within CRISPR leader regions that may regulate CRISPR adaptation through Cas6/RT/Cas1 systems and influence how RNA-DNA spacers are acquired and integrated. This computational work also informs wet-lab efforts to purify and characterize the Marinomonas mediterranea (MMB-1) adaptation complex, an example of a reverse transcriptase-Cas1 integrase fusion studied in vitro. We designed a consensus leader sequence for experimental validation and visualized conservation patterns of M. mediterranea’s Cas1 protein. By exploring how CRISPR leader sequences evolve across species, this research advances understanding of CRISPR adaptation and supports innovation in CRISPR-based biotechnology and genome engineering.