Yeast artificial chromosome transfer into human renal carcinoma cells by 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 carcinoma cell line by yeast spheroplast fusion and cationic lipids. While the latter approach proved inefficient, significant numbers of clones containing both vector arms were obtained by spheroplast fusion. FISH analyses 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.
Jülicher
K
K
Innere Klinik und Poliklinik (Tumorforschung), Universitatsklinikum Essen, Westdeutsches Tumorzentrum, Germany.
Vieten
L
L
Bröcker
F
F
Bardenheuer
W
W
Schütte
J
J
Opalka
B
B
D002292:Carcinoma, Renal Cell / Q000235:genetics*; D002459:Cell Fusion; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D002893:Chromosomes, Human, Pair 3 / Q000235:genetics; D003001:Cloning, Molecular; D018014:Gene Transfer Techniques; D016147:Genes, Tumor Suppressor; D005822:Genetic Vectors; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007680:Kidney Neoplasms / Q000235:genetics*; D013104:Spheroplasts; D014407:Tumor Cells, Cultured