The murine SP-C promoter directs type II cell-specific expression in transgenic mice.
Genomic DNA from the mouse pulmonary surfactant protein C (SP-C) gene was analyzed in transgenic mice to identify DNA essential for alveolar type II cell-specific expression. SP-C promoter constructs extending either 13 or 4.8 kb upstream of the transcription start site directed lung-specific expression of the bacterial chloramphenicol acetyl transferase (CAT) reporter gene. In situ hybridization analysis demonstrated alveolar cell-specific expression in the lungs of adult transgenic mice, and the pattern of 4.8 SP-C-CAT expression during development paralleled that of the endogenous SP-C gene. With the use of deletion constructs, lung-specific, low-level CAT activity was detected in tissue assays of SP-C-CAT transgenic mice retaining 318 bp of the promoter. In transient and stable cell transfection experiments, the 4.8-kb SP-C promoter was 90-fold more active as a stably integrated gene. These findings indicate that 1) the 4.8-kb SP-C promoter is sufficient to direct cell-specific and developmental expression, 2) an enhancer essential for lung-specific expression maps to the proximal 318-bp promoter, and 3) the activity of the 4.8-kb SP-C promoter construct is highly dependent on its chromatin environment.
Glasser
Stephan W
SW
Cincinnati Children's Hospital Medical Center, Division of Pulmonary Biology, 3333 Burnet Ave., Cincinnati, OH 45229-3039, USA. steve.glasser@cchmc.org
Eszterhas
Susan K
SK
Detmer
Emily A
EA
Maxfield
Melissa D
MD
Korfhagen
Thomas R
TR
D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011650:Pulmonary Alveoli / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D037721:Pulmonary Surfactant-Associated Protein C / Q000235:genetics* / Q000378:metabolism; D011663:Pulmonary Surfactants / Q000378:metabolism; D017384:Sequence Deletion; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D014162:Transfection
D002843:Chromatin; D037721:Pulmonary Surfactant-Associated Protein C; D011663:Pulmonary Surfactants; D015500:Chloramphenicol O-Acetyltransferase