In vivo selection method for determining inhibitory RNA molecules

摘要

A selection system suitable for use in vivo is provided, the system comprising: I) a plurality of first nucleotide sequences encoding a gene product capable of binding to and effecting the cleavage, directly or indirectly, of a nucleotide sequence, or a transcription product thereof; wherein a region of the first nucleotide sequence required for binding to the nucleotide sequence is heterogeneous within the plurality of first nucleotide sequences; and ii) a second nucleotide sequence comprising: (a) a coding region encoding a detectable marker operably linked to sequences required for mRNA stability and/or translation; and (b) a third nucleotide sequence positioned between the coding region and at least one of the sequences required for mRNA stability and/or translation; wherein (a) and (b) are operably linked to a regulatory sequence capable of directing expression of (a) and (b) as a contiguous RNA molecule in a host cell; and wherein the first nucleotide sequence encoding a gene product is capable of binding to and effecting the cleavage, directly or indirectly, of the third nucleotide sequence or a transcription product thereof.