Patent ID: 11866699
Assignee: UNIVERSITY OF WASHINGTON
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A composition comprising:
(a) a multimeric assembly, comprising a plurality of oligomeric substructures, wherein each oligomeric substructure comprises a plurality of recombinant polypeptides that self-interact around at least one axis of rotational symmetry, wherein
(i) each of the recombinant polypeptides comprises a polypeptide-polypeptide interface (“O interface”):
(ii) one or more of the recombinant polypeptides comprises a polypeptide domain that is capable of interacting with a lipid bilayer (“M domain”);
(iii) one or more of the recombinant polypeptides comprises a polypeptide domain that is capable of effecting membrane scission and release of an enveloped multimeric assembly from a cell by recruiting the ESCRT machinery to the site of budding by binding to one or more proteins in the eukaryotic ESCRT complex (“L domain”); and
(iv) one or more of the recombinant polypeptides comprises an RNA binding domain (“RBD”);

wherein the M domain, the L domain, the O interface, and the RBD are not each present in a single naturally occurring protein; and
(b) one or more active component selected from the group consisting of:
(i) one or more RNAs comprising
(A) one or more guide RNA (gRNA), wherein each gRNA comprises (I) a guide sequence capable of binding to a desired nucleic acid sequence; (II) an RNA packaging sequence capable of being bound by the RBD; and (III) a structural sequence capable of binding to a Class 2 Cas protein to form a Class 2 Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR)-Cas ribonucleoprotein (RNP) complex; and
(B) one or more mRNA encoding a Class 2 Cas protein and an RNA packaging sequence capable of being bound by the RBD; and/or

(ii) one or more Class 2 CRISPR-Cas RNP complexes comprising a Class 2 Cas protein and one or more gRNA bound by the Class 2 Cas protein, wherein each gRNA comprises (I) a guide sequence capable of binding to a desired nucleic acid sequence; (II) an RNA packaging sequence capable of being bound by the RBD; and (III) a structural sequence capable of binding to a Class 2 Cas protein;

wherein the structural sequence comprises or consists of a sequence that is at least 50% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NOS:332-335:, (SEQ ID NO: 332)

GUUUAAGAGCUA-N-UGCUGGAAACAGCAUAGCAAGUUUAAAUAAGGC

UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGC,

 

(SEQ ID NO: 333)

GUUUUAGUACUCUG-N-GAAACAGAAUCUACUAAAACAAGGCAAAAUG

CCGUGUUUAUCUCGUCAACUUGUUGGCGAGA,

 

(SEQ ID NO: 334)

AAUUUCUACUC-N-UUGUAGAU,

and

 

(SEQ ID NO: 335)

AAUUUCUACUAA-N-GUGUAGAU;, wherein N is absent or comprises or consists of an RNA packaging sequence.