YEAST ARTIFICIAL CHROMOSOME TRANSFER INTO HUMAN RENAL-CARCINOMA CELLSBY SPHEROPLAST FUSION

Successful transfer of yeast artificial chromosomes (YACs) into human cells has been described in only a single study. We here report on the evaluation of YAC transfer strategies into a human renal cell carcino ma cell Line by yeast spheroplast fusion and cationic lipids. While th e latter approach proved inefficient, significant numbers of clones co ntaining both vector arms were obtained by spheroplast fusion. FISH an alyses on such clones revealed the presence of YAC integration and the co-localization of both vector arms with insert sequences. These data demonstrate that under certain experimental conditions efficient YAC transfer into human cells by spheroplast fusion is possible and may be useful for the cloning of human disease-related genes. (C) 1997 Acade mic Press.