Methods and reagents for identifying synthetic genetic elements
OwnerJeno Gyurisinventor
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Abstract
The present invention relates to a selection method that allows fast recovery and identification of functional gene fragments which selectively inhibit growth, e.g., are cytostatic or cytotoxic, of particular cell-types, such as transformed cells. The strategy relies, in part, on the ability of small gene fragments to encode dominant-acting synthetic genetic elements (SGEs), e.g., molecules which interfere with the function of genes from which they are derived. SGEs which can be identified by the subject method include, but are not limited to, inhibitory antisense RNA molecules, ribozymes, nucleic acid decoys, and small peptides.
Claims22
A method for identifying agents which selectively confer a desired phenotype on a target cell, comprising:
- (i) transfecting subtractive cells with a library of expression vectors comprising a variegated population of coding sequences for potential synthetic genetic elements (SGEs);
- (ii) isolating, from the subtractive cells, those SGE vectors of the SGE library which do interfere with the detection of the desired phenotype in the target cell;
- (iii) transfecting target cells with the sub-population of SGE vectors isolated in step (ii);
- and
- (iv) isolating those SGE vectors which confer the desired phenotype to the target cell.
A method for identifying agents with selective antiproliferative activity for a target cell, comprising:
- (i) transfecting subtractive cells with a library of expression vectors comprising a variegated population of coding sequences for potential synthetic genetic elements (SGEs);
- (ii) isolating those SGE vectors of the SGE library that are not antiproliferative to the subtractive cells;
- (iii) transfecting target cells with the non-antiproliferative SGE vectors isolated in step (ii);
- and
- (iv) isolating those SGE vectors that are antiproliferative to the target cell.
Dependencies are inferred from the claim text.