Investigation of neural stem cell-specific regulatory promoter elements
- Authors:
- Honghua Yuan
- Ankang Hu
- Li Zhang
- Xiaorong Zhu
View Affiliations
Affiliations: Department of Neurobiology, Xuzhou Medical College, Xuzhou, Jiangsu 221002, P.R. China, Laboratory Animal Center, Xuzhou Medical College, Xuzhou, Jiangsu 221002, P.R. China
- Published online on: June 18, 2012 https://doi.org/10.3892/etm.2012.614
-
Pages:
405-408
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Abstract
The present study aimed to investigate neural stem cell (NSC)-specific regulatory promoter elements. PCR was employed to amplify the full sequence (4,000 bp) and core sequence (400 bp) of the promoter and intron-2 of the mouse nestin gene. pcDNA3.1 was used as a template to construct 6 different recombinant plasmids. CMV, CMV + intron-2, the full sequence of the nestin gene promoter, the full sequence of the nestin gene promoter + intron-2, the core sequence of the nestin gene promoter and the core sequence of the nestin gene promoter + intron-2 were independently used as promoters to regulate EGFP expression. The 6 recombinant plasmids were independently used to transfect nestin-positive and nestin-negative cells, and the expression of EGFP was observed under a fluorescence microscope. At the same time, flow cytometry was carried out to measure the proportion of cells positive for EGFP. The results showed that the full sequence and core sequence of the nestin gene promoter non-specifically regulated EGFP expression in cells and exhibit potent regulatory potency. The full sequence or core sequence of the nestin gene promoter which was fused with intron-2 can only regulate the EGFP expression in nestin-positive cells. CMV + intron-2 have non-specific regulation of EGFP alone. Thus, we conclude that the full sequence of the nestin gene promoter which is fused with intron-2 can specifically regulate the expression of exogenous genes in nestin-positive cells.
View References
1.
|
Reynolds BA and Weiss S: Generation of
neurons and astrocytes from isolated cells of the adult mammalian
central nervous system. Science. 255:1707–1710. 1992. View Article : Google Scholar : PubMed/NCBI
|
2.
|
Eriksson PS, Perfilieva E, Björk-Eriksson
T, Alborn AM, Nordborg C, Peterson DA and Gage FH: Neurogenesis in
the adult human hippocampus. Nat Med. 4:1313–1317. 1998. View Article : Google Scholar : PubMed/NCBI
|
3.
|
Nakano T: Establishment maintenance and
differentiation induction of embryonic stem cells. Nihon Rinsho.
61:385–389. 2003.PubMed/NCBI
|
4.
|
Cheng L, Jin Z, Liu L, Yan Y, Li T, Zhu X
and Jing N: Characterization and promoter analysis of the mouse
nestin gene. FEBS Lett. 565:195–202. 2004. View Article : Google Scholar : PubMed/NCBI
|
5.
|
Lothian C and Lendahl U: An evolutionarily
conserved region in the second intron of the human nestin gene
directs gene expression to CNS progenitor cells and to early neural
crest cells. Eur J Neurosci. 9:452–462. 1997. View Article : Google Scholar
|
6.
|
Trinklein ND, Aledred SJ, Saldanha AJ and
Myers RM: Identification and functional analysis of human
transcriptional promoters. Genome Res. 13:308–312. 2003. View Article : Google Scholar : PubMed/NCBI
|
7.
|
Sun HX, Lu DX and Liu FP: Theory and
Application of Transgenic Technology Henan Medical. University
Press; Zhengzhou: pp. 2342000
|
8.
|
Diamond MI, Miner JN, Yoshinaga SK and
Yamamoto KR: Transcription factor interactions: selectors of
positive or negative regulation from a single DNA element. Science.
249:1266–1272. 1990. View Article : Google Scholar : PubMed/NCBI
|
9.
|
Josephson R, Müller T, Pickel J, Okabe S,
Reynolds K, Turner PA, Zimmer A and McKay RD: POU transcription
factors control expression of CNS stem cell-specific genes.
Development. 125:3087–3100. 1998.PubMed/NCBI
|
10.
|
Tanaka S, Kamachi Y, Tanouchi A, Hamada H,
Jing N and Kondoh H: Interplay of SOX and POU factors in regulation
of the nestin gene in neural primordial cells. Mol Cell Biol.
24:8834–8846. 2004. View Article : Google Scholar : PubMed/NCBI
|