Genomic sequences capable of committing mouse and rat fibroblasts to adipogenesis.
The mouse Swiss 3T3-F442A/3T3-C2 cell system is well suited for the isolation of genes involved in commitment to adipogenesis. 3T3-F442A cells convert to adipocytes with high efficiency in response to confluence and insulin. The sister clonal line 3T3-C2 does not respond to these signals, but can convert to adipocytes when transfected with DNA from 3T3-F442A preadipocytes or from human fat. Human fat-tissue biopsy FO46 DNA transfected into 3T3-C2 gave rise to fat foci after two rounds of transfection and selection. A cosmid library of a subclone of secondary transfectant 3T3-C2/FO46-1 was screened for the human repetitive Alu sequence. Five out of eight Alu+ recombinant clones committed 3T3-C2 cells to adipogenesis. The adipose commitment (AC) activity of one cosmid, p18A4, was found to reside in two small, non-identical, subcloned sequences 1.2kb and 2.0kb in length, each separately able to commit 3T3-C2, precrisis mouse and rat fibroblasts and the multipotential C3H10T1/2 cell line to adipogenesis. We conclude that commitment to adipogenesis can be effected in vitro with high efficiency by transfection of specific sequences into a variety of host cells.
Colón-Teicher
L
L
Department of Biological Sciences, Columbia University, New York, NY 10027.
Wise
L S
LS
Martino
J J
JJ
Baskin
L
L
Sakoulas
G
G
Pollack
R E
RE
Chen
S
S
D016475:3T3 Cells; D000273:Adipose Tissue / Q000254:growth & development*; D000818:Animals; D015139:Blotting, Southern; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D003360:Cosmids; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D014162:Transfection