Inactivating expression of selected protein or RNA in cells comprises integrating a sequence into transcript of selected protein or RNA which acts as target for interfering RNA

摘要

Inactivating expression of a selected protein or RNA in a cell, in vitro or ex vivo comprises: (A) introducing into the cell a vector (V1) that contains (i) a nucleic acid (A) encoding the protein or RNA and (ii) at least two copies of a sequence (B) of at least 10 nucleotides, so that during transcription, B makes up part of the transcript containing A but does not make up part of the coding sequence comprising A; and (B) introducing into the cell a second vector (V2) that causes synthesis of an interfering RNA (iRNA) that targets B. Independent claims are also included for the following: (1) combination of V1 and V2; (2) vector (V3) comprising a multiple cloning site for insertion of A and at least 2 copies of B; (3) vector (V4) containing A and at least two copies of B so that during transcription B makes up part of the 3'-UTR (untranslated region) of the A-containing transcript; (4) viral particles comprising V3 or V4, where these are viral vectors; (5) kit for expressing a protein or RNA, and for its inactivation, comprising V1 and V2; and (6) genetically modified cell containing in its genome an exogenous transgene that includes, in its 3'-UTR, at least two copies of B, optionally also a sequence that produces iRNA.