Patent ID: 6268134
Filing Date: 2001-07-31
Classification: C12N,C12Q,C40B

Abstract:
A method for producing GSEs corresponding to recessive genes that, when inactivated by GSEs, confer a selectable or screenable phenotype upon a cell having such inactive genes, the method comprising the steps of:(a) obtaining genomic DNA or a total cDNA population from the cells;(b) randomly fragmenting the genomic DNA or total cDNA population to produce random DNA fragments;(c) ligating the random DNA fragments to synthetic adaptors to produce amplifiable random DNA fragments;(d) cloning the amplified mixture of random DNA fragments into a suitable expression vector having a selectable marker to produce a random fragment expression library;(e) transferring the random fragment expression library into appropriate the random fragment expression library into appropriate target cells;(f) selecting the target cells for the presence of the selectable marker present in the expression vector to obtain target cells having the selectable marker;(g) selecting or screening the target cells having the selectable marker for the selectable or screenable phenotype conferred upon the cells by inactivation of a recessive gene by a GSE;(h) recovering the GSE from the target cell having the selectable or screenable phenotype.