Effect of 5'-flanking sequence deletions on expression of the human insulin gene in transgenic mice
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Effect of 5'-flanking sequence deletions on expression of the human insulin gene in transgenic mice. / Fromont-Racine, M; Bucchini, D; Madsen, O; Desbois, P; Linde, S; Saulnier, C; Ripoche, M A; Jami, J; Pictet, R; Nielsen, Jens Høiriis.
I: Molecular endocrinology (Baltimore, Md.), Bind 4, Nr. 5, 05.1990, s. 669-77.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - Effect of 5'-flanking sequence deletions on expression of the human insulin gene in transgenic mice
AU - Fromont-Racine, M
AU - Bucchini, D
AU - Madsen, O
AU - Desbois, P
AU - Linde, S
AU - Saulnier, C
AU - Ripoche, M A
AU - Jami, J
AU - Pictet, R
AU - Nielsen, Jens Høiriis
PY - 1990/5
Y1 - 1990/5
N2 - Expression of the human insulin gene was examined in transgenic mouse lines carrying the gene with various lengths of DNA sequences 5' to the transcription start site (+1). Expression of the transgene was demonstrated by 1) the presence of human C-peptide in urine, 2) the presence of specific transcripts in pancreas, but not in other tissues, 3) the specific immunofluorescence staining of pancreatic islets for human C-peptide, and 4) the synthesis and accumulation of human (pro)insulin in isolated islets. Deletions in the injected DNA fragment of sequences upstream from positions -353, -258, and -168 allowed correct initiation of the transcripts and cell specificity of expression, while quantitative expression gradually decreased. Deletion to -58 completely abolished the expression of the gene. The amount of human product that in mice harboring the longest fragment contributes up to 50% of the total insulin does not alter the normal proportion of mice insulins I and II. These results suggest that expression of the human insulin gene in vivo results from the cooperation of several cis-regulatory elements present in the various deleted fragments. With none of the deletions used, expression of the transgene was observed in cell types other than beta-islet cells.
AB - Expression of the human insulin gene was examined in transgenic mouse lines carrying the gene with various lengths of DNA sequences 5' to the transcription start site (+1). Expression of the transgene was demonstrated by 1) the presence of human C-peptide in urine, 2) the presence of specific transcripts in pancreas, but not in other tissues, 3) the specific immunofluorescence staining of pancreatic islets for human C-peptide, and 4) the synthesis and accumulation of human (pro)insulin in isolated islets. Deletions in the injected DNA fragment of sequences upstream from positions -353, -258, and -168 allowed correct initiation of the transcripts and cell specificity of expression, while quantitative expression gradually decreased. Deletion to -58 completely abolished the expression of the gene. The amount of human product that in mice harboring the longest fragment contributes up to 50% of the total insulin does not alter the normal proportion of mice insulins I and II. These results suggest that expression of the human insulin gene in vivo results from the cooperation of several cis-regulatory elements present in the various deleted fragments. With none of the deletions used, expression of the transgene was observed in cell types other than beta-islet cells.
KW - Animals
KW - C-Peptide
KW - Chromosome Deletion
KW - Female
KW - Gene Expression
KW - Humans
KW - Immunohistochemistry
KW - Insulin
KW - Male
KW - Mice
KW - Mice, Transgenic
KW - Pancreas
KW - RNA, Messenger
M3 - Journal article
C2 - 2274051
VL - 4
SP - 669
EP - 677
JO - Molecular Endocrinology
JF - Molecular Endocrinology
SN - 0888-8809
IS - 5
ER -
ID: 47974095