[3S03m] 08. Functional Molecules and Materials for Biological Systems - Part 1

10:40 AM - 11:20 AM JST(1:40 AM - 2:20 AM UTC)

[3FMBS-03] Programmable Gene Editing and RNA Delivery for Regenerative Medicine and Sustainable Biotechnology

○Yu-Chen Andy Hu¹ (1. National Tsing Hua University (Taiwan))

Keywords:

CRISPR,tissue regeneration,RNA editing,RNA Delivery,gene regulation

Recent advances in gene editing and gene regulation technologies have greatly expanded our ability to precisely manipulate cellular functions and engineer biological systems. We have developed several CRISPR-based gene regulation platforms, including CRISPR activation (CRISPRa), CRISPR inhibition (CRISPRi), and bi-directional gene regulation systems that enable simultaneous gene activation and repression. These technologies provide powerful tools for controlling cellular signaling pathways and have been applied to guide stem cell differentiation and enhance bone regeneration. In addition, we have developed RNA editing strategies for microRNA knockdown to further expand the toolbox for stem cell engineering and regenerative medicine. Furthermore, efficient delivery of genetic cargos remains a major challenge for gene therapy and RNA therapeutics. To address this issue, we developed a novel protein-based nanoparticle platform derived from the human retroelement PEG10, which enables spontaneous self-packaging of cargo RNA and efficient delivery into target cells. This platform provides a promising strategy for RNA delivery and cancer therapy.Beyond biomedical applications, gene editing technologies can also be applied to engineer microbial systems for sustainable biomanufacturing. Using CRISPR-based genome engineering and regulatory tools, we engineered several non-model industrial microorganisms—including cyanobacteria, Pseudomonas putida, and the yeast Candida viswanathii—to produce a variety of bio-based chemicals. In particular, metabolic engineering of Candida viswanathii enabled high-level production of C12 dicarboxylic acids, achieving titers exceeding 369 g/L in fermenters. We further implemented an RNA-level regulatory system based on CRISPR-Cas13d (hfCas13d) with circular guide RNAs to achieve sustained and multiplex pathway regulation. Multiplex regulation using dual guide RNA arrays improved 10-hydroxydecanoic acid production to 37.2 g/L. Together, these studies highlight how advances in gene editing, gene delivery, and RNA delivery technologies can enable new opportunities in regenerative medicine, RNA therapeutics, and sustainable biotechnology.