Engineered MCP and PCP proteins and systems and methods thereof
AssigneeTrustees of Boston University
InventorsJohn T. Ngo, Christopher Joseph Kuffner, Alex Marzilli
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Abstract
The technology described herein is directed to engineered MCP proteins and engineered PCP proteins, which are degraded when the proteins are not bound to an MS2 or PP7 RNA hairpin loop, respectively. Also described herein are fusion proteins comprising such engineered MCP proteins and engineered PCP proteins linked to various effector proteins. The linkage to the effector proteins can be modulated through of specialized linker domains. In addition, described herein are complexes and systems comprising the fusion proteins in combination with synthetic RNA molecules, in order to modulate the structure and/or function the synthetic RNA molecules.
Claims27
An engineered MS2 coat protein (MCP) comprising an engineered RNA-binding domain;
- wherein the engineered MCP protein is stable when the engineered RNA-binding domain is bound to an MS2 RNA hairpin loop;
- wherein when the engineered RNA-binding domain is not bound to the MS2 RNA hairpin loop, the engineered MCP protein is unstable and degradation of the engineered MCP protein increases;
- wherein the engineered RNA-binding domain comprises:
- (a) a C-terminal portion of a first MCP monomer;
- and
- (b) an N-terminal portion of the first MCP monomer;
- wherein the first MCP monomer is split into the N-terminal portion and the C-terminal portion at a binding pocket for the MS2 RNA hairpin loop.
An engineered MS2 coat protein (MCP) comprising an engineered RNA-binding domain and a degron;
- wherein the degron is hidden upon binding of the engineered RNA-binding domain to an MS2 RNA hairpin loop;
- wherein when the engineered RNA-binding domain is not bound to the MS2 RNA hairpin loop, the degron is exposed and degradation of the engineered MCP protein increases;
- wherein the engineered RNA-binding domain comprises:
- (a) a C-terminal portion of a first MCP monomer;
- and
- (b) an N-terminal portion of the first MCP monomer;
- wherein the first MCP monomer is split into the N-terminal portion and the C-terminal portion at a binding pocket for the MS2 RNA hairpin loop.
Dependencies are inferred from the claim text.