Characterization of five evolutionary conserved regions of the human tyrosine hydroxylase (TH) promoter: implications for the engineering of a human TH minimal promoter assembled in a self-inactivating lentiviral vector system.
A DNA fragment of about 13 kb containing the human tyrosine hydroxylase (TH) promoter was previously isolated from a genomic DNA library and sequenced. The 11 kb from the transcription start of the human TH promoter was successively joined to the green fluorescent protein (GFP) to generate a transgenic mouse model. High levels of GFP expression could be observed in TH-positive cells of the Substantia nigra of embryonic and adult mice. Intriguingly, the sequence of the human TH promoter showed a low degree of homology with the mouse and rat TH promoters. In fact, comparative analysis of the sequences of human, rat, and mouse TH promoters revealed only five small regions of high homology. These five evolutionarily conserved regions were numbered in numeric progression from the 5' end of human TH promoter. In the present study, a panel of minimal human TH promoters was generated to analyze the transcriptional activity and specificity of gene expression conferred by the five conserved regions (CRs). The series of constructs was termed 250 bp and contained the first -194 bp of the human TH promoter immediately upstream of the transcription start, the first 35 bp the human TH messenger RNA leader, plus one or more of the five CRs. All the constructs were assembled in a self-inactivating form of the latest series of lentiviral vector system based on the human immunodeficiency virus type 1 (HIV-1). Lentiviral-mediated gene transfer was highly efficient for the in vitro transduction of human neuronal progenitor cells (hNPCs). Since a subset of hNPCs express TH following in vitro treatment with a mixture of differentiating agents, it was possible to assess specificity of expression for all the minimal human TH promoters. Overall, the successive addition of the five conserved regions produced a greater degree of specificity in induced TH-positive hNPCs, in particular after the addition of CRI (-8,917, -8,876). However, the human TH minimal promoters did not show any specificity for TH-positive differentiated mouse primary striatal and S. nigra cells, indicating a difference of TH gene regulation between the human and mouse systems. The human TH minimal promoters may provide the opportunity for the selection of TH-positive human embryonic and adult stem cells for brain transplantation experiments in animal models for Parkinson's disease.
Romano
Gaetano
G
Department of Neurosurgery, The Farber Institute for the Neurosciences, Thomas Jefferson University, Jefferson Hospital for the Neurosciences, Philadelphia, Pennsylvania, USA. Gaetano.Romano@jefferson.edu
Suon
Sokreine
S
Jin
Hao
H
Donaldson
Angela E
AE
Iacovitti
Lorraine
L
D000818:Animals; D001698:Biomedical Engineering; D002478:Cells, Cultured; D017124:Conserved Sequence; D003342:Corpus Striatum / Q000166:cytology / Q000378:metabolism; D019143:Evolution, Molecular; D020869:Gene Expression Profiling; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D051381:Rats; D013378:Substantia Nigra / Q000166:cytology / Q000378:metabolism; D013379:Substrate Specificity; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics* / Q000378:metabolism
D014446:Tyrosine 3-Monooxygenase
Neurobiol Dis. 2004 Aug;16(3):596-603
15262272
FEBS Lett. 1986 Aug 11;204(1):5-7
3091398
Annu Rev Neurosci. 1989;12:415-61
2564757
Nucleic Acids Res. 1990 May 25;18(10):3093
2349127
J Neurochem. 1990 Dec;55(6):2149-52
1977891
Biochemistry. 1993 Aug 24;32(33):8397-402
8357791
Biochem Biophys Res Commun. 2003 Dec 26;312(4):1123-31
14651989
Science. 1994 Aug 26;265(5176):1221-5
8066460
Nucleic Acids Res. 1995 Feb 25;23(4):628-33
7899083
Int Rev Cytol. 1995;162A:279-388
8575883
Science. 1996 Apr 12;272(5259):263-7
8602510
J Neurochem. 1997 Feb;68(2):564-9
9003041
Science. 1997 Apr 11;276(5310):248-50
9092472
J Neurochem. 1997 May;68(5):1792-803
9109503
J Neurochem. 1997 May;68(5):2152-60
9109543
Hum Mol Genet. 1998 Mar;7(3):423-8
9466999
Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):4013-8
9520484
Mol Cell Neurosci. 1998 May;11(1-2):36-46
9608532
Hum Gene Ther. 1999 Jul 20;10(11):1763-71
10446916
Neuroscience. 1999;93(2):631-42
10465447
J Biol Chem. 1964 Sep;239:2910-7
14216443
Science. 2002 Feb 1;295(5556):868-72
11786607
Brain Res Dev Brain Res. 2001 Sep 23;130(1):91-8
11557097
Hum Mol Genet. 2001 Aug 15;10(17):1785-92
11532988
Biochem Biophys Res Commun. 2000 Aug 11;274(3):590-5
10924322
J Neurochem. 2000 May;74(5):1829-37
10800925
Stem Cells. 2000;18(1):19-39
10661569
Nat Neurosci. 2000 Jan;3(1):41-6
10607393
Nat Med. 1999 Nov;5(11):1256-63
10545991
Nat Med. 1999 Nov;5(11):1237-8
10545984
Neurosci Lett. 2004 Jun 3;363(1):33-7
15157991
Eur J Dermatol. 2003 Sep-Oct;13(5):424-9
14693483
Cell Transplant. 2004;13(5):535-47
15565866
Acta Biochim Pol. 2003;50(3):865-73
14515167
Drug News Perspect. 2003 Jun;16(5):267-76
12942157
J Neurochem. 2003 May;85(3):622-34
12694388
Brain Res Mol Brain Res. 2003 Apr 10;112(1-2):8-23
12670698
Neurosci Lett. 2002 Sep 20;330(2):151-4
12231434
Nature. 2002 Jul 4;418(6893):50-6
12077607
Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8392-6
7690960