Unravelling the genetic codes governing transposon mobilization

Transposons are ubiquitous DNAs capable of changing their genomic locations. While historically perceived as detrimental and damaging to host genomes, they have also been a major source of genetic variability, driving evolution. Unfortunately, the regulatory mechanisms of transposons remain largely unknown, primarily due to the lack of experimental methods to trace their movement. This project therefore aims to dissect the regulation of transposons in plants with unprecedented detail. This goal is enabled by a distinctive method developed in my lab, RUM (Real-time detection of transposon jUMping). RUM provides real-time, single-cell detection of transposon movement, offering a powerful platform to unravel the complexities of transposition regulation. Specifically, this project will focus on three major objectives investigating at multiple scales, from tissue to cellular and molecular levels. Firstly, it has long been assumed that transposition occurs more frequently in somatic tissues, however, this assumption has yet to be empirically validated. To comprehensively evaluate cell type-specific transposition activity, I will integrate RUM with single-cell RNA sequencing. This approach will provide a detailed map of cellular competence for transposition. Secondly, while epigenetic silencing is well known as a mechanism controlling transposons, significant gaps remain in our understanding of post-transcriptional and pre-integrational regulation. To identify novel regulators of transposition, I will perform a custom genome-wide CRISPR screen in combination with RUM. Thirdly, internal and auto-regulatory sequence motifs within transposons remain poorly studied. To address this, I will generate random mutations within RUM and map mutation hotspots associated with active transposition. Altogether, this study will illuminate the complex regulatory networks governing transposon movement, bridging longstanding knowledge gaps in biology.