Transgene expression in the QM myogenic cell line.
We have isolated an avian muscle cell line (QM) which has the essential features of established mammalian muscle cell lines. The experiments reported here were undertaken to determine the suitability of QM cells for the introduction and analysis of cloned transgenes. The promoter of the cardiac troponin T (cTNT) gene has been previously shown to contain sequence elements which govern muscle-specific expression of the chloramphenicol acetyltransferase (CAT) gene in transiently transfected primary cell cultures. We show here that QM cells stably harboring cTNT promoter-CAT fusion genes up-regulate CAT expression in concert with myogenic differentiation, and that as few as 110 upstream nucleotides are sufficient for such differentiation-dependent regulation. In addition, both transient and stable transfection experiments demonstrate that differentiated QM cells possess trans-acting factors necessary for the expression of the skeletal alpha-actin promoter, despite the absence of mRNA or protein product from the endogenous sarcomeric actin genes in these cells. Finally, to follow the developmental potential of QM cells in vivo, we created a clone, QM2ADH, which constitutively expresses the histochemical marker transgene encoding Drosophila alcohol dehydrogenase. When surgically inserted into the limb buds of developing chick embryos, QM2ADH cells are incorporated into endogenous developing muscles, indicating that QM cells are capable of recognizing and responding to host cues governing muscle morphogenesis. Thus, QM cells are versatile as recipients of transgenes for the in vitro and in vivo analysis of molecular events in muscle development.
Antin
P B
PB
Department of Anatomy, University of California, San Francisco 94143.
Karp
G C
GC
Ordahl
C P
CP
D000199:Actins / Q000235:genetics*; D000426:Alcohol Dehydrogenase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular; D015870:Gene Expression; D007700:Kinetics; D009024:Morphogenesis; D009132:Muscles / Q000166:cytology / Q000196:embryology / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011784:Quail; D014162:Transfection; D014336:Troponin / Q000235:genetics*; D020107:Troponin T; D015854:Up-Regulation
D000199:Actins; D014336:Troponin; D020107:Troponin T; D000426:Alcohol Dehydrogenase; D015500:Chloramphenicol O-Acetyltransferase