Contact

  • Academic
    kangjw@stanford.edu
    University - Student Department: School of Medicine - IDP's - Neurosciences Position: Graduate

Additional Info

  • Mail Code: 5420

All Publications

  • Tunable, self-contained gene dosage control via proteolytic cleavage of CRISPR-Cas systems. bioRxiv : the preprint server for biology Katz, N., An, C., Lee, Y. J., Tycko, J., Zhang, M., Kang, J., Bintu, L., Bassik, M. C., Huang, W. H., Gao, X. J. 2024

    Abstract

    Gene therapy holds great therapeutic potential. Yet, controlling cargo expression in single cells is limited due to the variability of delivery methods. We implement an incoherent feedforward loop based on proteolytic cleavage of CRISPR-Cas activation or inhibition systems to reduce gene expression variability against the variability of vector delivery. We demonstrate dosage control for activation and inhibition, post-delivery tuning, and RNA-based delivery, for a genome-integrated marker. We then target the RAI1 gene, the haploinsufficiency and triplosensitivity of which cause two autism-related syndromes, Smith-Magenis-Syndrome (SMS) and Potocki-Lupski-Syndrome, respectively. We demonstrate dosage control for RAI1 activation in HEK293s, Neuro-2As, and mouse cortical neurons via AAVs and lentiviruses. Finally, we activate the intact RAI1 copy in SMS patient-derived cells to an estimated two-copy healthy range, avoiding the harmful three-copy regime. Our circuit paves the way for viable therapy in dosage-sensitive disorders, creating precise and tunable gene regulation systems for basic and translational research.

    View details for DOI 10.1101/2024.10.09.617463

    View details for PubMedID 39416069

    View details for PubMedCentralID PMC11482798

  • Protease-controlled secretion and display of intercellular signals. Nature communications Vlahos, A. E., Kang, J., Aldrete, C. A., Zhu, R., Chong, L. S., Elowitz, M. B., Gao, X. J. 2022; 13 (1): 912

    Abstract

    To program intercellular communication for biomedicine, it is crucial to regulate the secretion and surface display of signaling proteins. If such regulations are at the protein level, there are additional advantages, including compact delivery and direct interactions with endogenous signaling pathways. Here we create a modular, generalizable design called Retained Endoplasmic Cleavable Secretion (RELEASE), with engineered proteins retained in the endoplasmic reticulum and displayed/secreted in response to specific proteases. The design allows functional regulation of multiple synthetic and natural proteins by synthetic protease circuits to realize diverse signal processing capabilities, including logic operation and threshold tuning. By linking RELEASE to additional sensing and processing circuits, we can achieve elevated protein secretion in response to "undruggable" oncogene KRAS mutants. RELEASE should enable the local, programmable delivery of intercellular cues for a broad variety of fields such as neurobiology, cancer immunotherapy and cell transplantation.

    View details for DOI 10.1038/s41467-022-28623-y

    View details for PubMedID 35177637