The rapid pace of innovation in synthetic biology and genome engineering elicits a need to reevaluate systems of oversight to ensure that biosafety and biosecurity safeguards are keeping up. Accordingly, the regulation of nucleic acid synthesis, an enabling technology for synthetic biology and genome engineering, is a current focus of political debate in the United States. However, to develop appropriate governance that neither under- nor overregulates technological development, policy leaders must also appreciate how advances in synthetic biology and genome engineering are being employed in the interests of biosecurity to support human health, manufacturing, food security, ecosystems and the natural environment. Synthetic biology can support the development of new therapeutics and health technologies, alternative biomanufacturing methodologies, food and agricultural innovations, environmental biosensors for monitoring infectious and toxic agents, and interventions aimed at preserving and restoring our natural environment. This Perspective provides an overview of current and potential benefits of synthetic biology and genome engineering, aiming to balance the broader societal discussion of potential risks—particularly in this special issue of the journal—with potential social, economic, and environmental value to individuals and society at large.
C. Chapman, Julie Trolle, Dominika Wawrzyniak et al.· Frontiers in Bioengineering...· 0 citations
Hirschsprung disease (HSCR) is a complex developmental disorder of the enteric nervous system, primarily driven by regulatory variants within enhancer elements of the RET gene. To investigate how these variants lead to aganglionosis, we developed a humanized mouse model by inserting an intact 77kb human RET genomic locus into the Rosa26 safe-harbor locus. Utilizing “big DNA” synthetic biology and Bxb1-mediated recombination, we integrated the complete human locus including all exons, introns, and upstream regulatory elements which we validated via nanopore and short-read sequencing. Functional analysis confirmed in vivo human RET expression; however, our initial HSCR-associated “sensitive” haplotype expressed at only 21% of wild-type levels. This significant reduction proved insufficient to rescue the viability when endogenous mouse Ret was deleted. We identified that this deficiency is partially driven by five risk SNPs within established enhancers. Specifically, using CRISPR/Cas9 to restore a conserved So×10 binding site (converting a sensitive SNP to a protective one) increased RET expression by 1.9-fold and restored transcription factor binding. This study provides a robust framework for modeling human-specific regulatory disorders and demonstrates the critical impact of non-coding variation on disease pathogenesis.
Ryan D. Fine, B. Low, Jarod A. Rollins et al.· bioRxiv· 0 citations