Year: 2023
Type: article
Abstract: Adeno-associated virus (AAV)-based gene therapy could be facilitated by the development of molecular switches to control the magnitude and timing of expression of therapeutic transgenes. RNA interference (RNAi)-based approaches hold unique potential as a clinically proven modality to pharmacologically regulate AAV gene dosage in a sequence-specific manner. We present a generalizable RNAi-based rheostat wherein hepatocyte-directed AAV transgene expression is silenced using the clinically validated modality of chemically modified small interfering RNA (siRNA) conjugates or vectorized co-expression of short hairpin RNA (shRNA). For transgene induction, we employ REVERSIR technology, a synthetic high-affinity oligonucleotide complementary to the siRNA or shRNA guide strand to reverse RNAi activity and rapidly recover transgene expression. For potential clinical development, we report potent and specific siRNA sequences that may allow selective regulation of transgenes while minimizing unintended off-target effects. Our results establish a conceptual framework for RNAi-based regulatory switches with potential for infrequent dosing in clinical settings to dynamically modulate expression of virally-delivered gene therapies.
Source: Nature Communications
Authors: Megha Subramanian, James McIninch, Ivan Zlatev, Mark K. Schlegel, Charalambos Kaittanis, Tuyen Nguyen, Saket Agarwal, Timothy Racie, Martha Arbaiza Alvarado, Kelly Wassarman, Thomas S. Collins, Tyler Chickering, Christopher R. Brown, Karyn Schmidt, Adam B. Castoreno, Svetlana Shulga-Morskaya, Elena Stamenova, Kira Buckowing, Daniel Berman, Joseph D. Barry, Anna Bisbe, Martin A. Maier, Kevin Fitzgerald, Vasant Jadhav
Institution: Alnylam Pharmaceuticals (United States)
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $6,290
Funder: Alnylam Pharmaceuticals
OpenAlex ID: W4362716453