Molecular mechanisms, functions and applications of RNA-guided DNA transposons
Genome engineering has transformed life science research and enabled revolutionary applications in biotechnology and medicine. Despite recent advances brought by the development of CRISPR-associated genome editing nucleases, a uni...
Genome engineering has transformed life science research and enabled revolutionary applications in biotechnology and medicine. Despite recent advances brought by the development of CRISPR-associated genome editing nucleases, a universal strategy for efficient site-specific gene editing and insertion in a range of organisms or cell types is currently unavailable. Transposons are natural genome remodelers and gene delivery vectors that modify and transfer genetic information across the tree of life by moving within and between their host genomes. To this end, they encode a vast and diverse reservoir of nucleic acid-targeting and -processing enzymes, whose molecular mechanisms are yet mostly unexplored. Building on our recent structural and biochemical studies, the goal of this project is to advance our mechanistic knowledge of RNA-guided DNA transposons in order to understand their biology and guide their repurposing as next-generation genetic tools. Specifically, the proposed studies will investigate (i) the mechanisms of RNA-directed DNA mobilization of CRISPR-associated transposons (CASTs), aiming not only to provide insights into their molecular function but also to inform their technological development, (ii) the molecular activities and architectures of TnpB-like transposon proteins, recently identified putative RNA-guided endonucleases of prokaryotic and eukaryotic origin, with the ultimate goal of supporting their application as ultra-compact, broadly applicable genome editors, and (iii) the translation of designer CASTs into a novel programmable DNA insertion technology for precision genomic profiling and engineering. Altogether, the proposed studies will not only make key contributions to the molecular understanding and harnessing of RNA-guided transposons in nature and technologies, but hold the long-term potential to address current challenges in genome editing, biotechnologies and precision medicine.ver más
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