	US 11,946,066 B2
	RNA-based delivery systems with levels of control
Xiaojing Gao, Pasadena, CA (US); Lucy S. Chong, Pasadena, CA (US); Michael B. Elowitz, Pasadena, CA (US); and Matthew S-M Kim, San Francisco, CA (US)
Assigned to California Institute of Technology, Pasadena, CA (US)
Filed by California Institute of Technology, Pasadena, CA (US)
Filed on Sep. 23, 2022, as Appl. No. 17/935,024.
Application 17/935,024 is a continuation of application No. 16/555,604, filed on Aug. 29, 2019, granted, now 11,453,893.
Claims priority of provisional application 62/777,420, filed on Dec. 10, 2018.
Claims priority of provisional application 62/725,020, filed on Aug. 30, 2018.
Prior Publication US 2023/0193317 A1, Jun. 22, 2023
Int. Cl. C12N 15/86 (2006.01); C07K 14/005 (2006.01); C07K 14/81 (2006.01); C07K 16/18 (2006.01); C12N 7/00 (2006.01); C12N 9/50 (2006.01); C12N 15/62 (2006.01)

CPC C12N 15/86 (2013.01) [C07K 14/005 (2013.01); C07K 16/18 (2013.01); C12N 7/00 (2013.01); C12N 9/506 (2013.01); C12N 15/62 (2013.01); C12Y 304/21098 (2013.01); C07K 2317/569 (2013.01); C07K 2317/622 (2013.01); C07K 2319/30 (2013.01); C07K 2319/50 (2013.01); C12N 2740/11045 (2013.01); C12N 2760/20143 (2013.01); C12N 2810/6054 (2013.01); C12N 2820/002 (2013.01); C12N 2820/007 (2013.01)]

20 Claims

1. A viral vector, comprising:

a polynucleotide encoding a nucleoprotein (N), a phosphoprotein (P), a matrix protein (M), an RNA-dependent RNA polymerase (L), and one or more transgenes,

wherein at least one of the N, P, M, or L is a degron fusion protein comprising a degron capable of binding a degron stabilizing molecule, and

wherein the degron fusion protein changes from a destabilized state to a stabilized state when the degron binds to the degron stabilizing molecule;

wherein at least one of the N, P, M, or L is a conditionally stable fusion protein comprising a stabilizing molecule binding domain capable of binding a stabilizing molecule, and wherein the conditionally stable fusion protein changes from a destabilized state to a stabilized state when the stabilizing molecule binding domain binds to the stabilizing molecule, and/or

wherein two of the N, P, M, or L are expressed as a protease fusion protein comprising a first protein and a second protein of the two of the N, P, M, or L separated from a protease by a first cut site and a second cut site, respectively, wherein the protease is capable of cutting the first cut site and the second cut site when the protease is not bound by a protease inhibitor, and wherein the first protein and the second protein are in inactive states when the first cut site and the second cut site are not cut, respectively.