Short interfering RNAs (siRNAs) for reducing dopaminergic phenotypic markers

被引：11

作者：

Bäckman, C ^{[1
]}

Zhang, YJ ^{[1
]}

Hoffer, BJ ^{[1
]}

Tomas, AC ^{[1
]}

机构：

[1] NIDA, Cellular Neurobiol Branch, NIH, Baltimore, MD 21224 USA

来源：

JOURNAL OF NEUROSCIENCE METHODS | 2003年 / 131卷 / 1-2期

基金：

美国国家卫生研究院;

关键词：

RNA interference; RNAi; short interfering RNA; siRNA; dopamine; midbrain; Nurr1; cRet; tyrosine hydroxylase;

D O I：

10.1016/S0165-0270(03)00236-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Short interfering RNAs (siRNAs) are double-stranded RNAs of similar to19-29 nucleotides designed to suppress the expression of homologous genes by a process known as RNA interference (RNAi). In this study we have characterized several short interfering RNAs to reduce (knockdown) the expression of genes related to the dopaminergic system of the ventral mesencephalon. We report here effective suppression of all targeted genes, tyrosine hydroxylase (TH), the transcription factor Nr4a2 (Nurr1) and the GDNF receptor moiety cRet, by co-transfection of plasmids expressing gene specific siRNAs under control of the human U6 promoter with a reporter plasmid coding for firefly luciferase and fused in tandem to the cDNA for the gene of interest. The effective suppression of Nr4a2, cRet and tyrosine hydroxylase suggest that the U6 expression cassette could be used to deliver siRNA to mammalian cells in vivo and actively suppress the expression of dopamine related mammalian genes. The characterization of highly effective siRNAs for DA phenotypic markers may open new avenues for loss-of-function phenotypic analysis, and may lead to new approaches for gene therapy. (C) 2003 Elsevier B.V. All rights reserved.

引用

页码：51 / 56

页数：6

共 24 条

[1] A selective group of dopaminergic neurons express Nurr1 in the adult mouse brain
Bäckman, C
Perlmann, T
Wallén, Å
Hoffer, BJ
Morales, M
[J]. BRAIN RESEARCH, 1999, 851 (1-2) : 125 - 132
[2] Role for a bidentate ribonuclease in the initiation step of RNA interference
Bernstein, E
Caudy, AA
Hammond, SM
Hannon, GJ
[J]. NATURE, 2001, 409 (6818) : 363 - 366
[3] A system for stable expression of short interfering RNAs in mammalian cells
Brummelkamp, TR
Bernards, R
Agami, R
[J]. SCIENCE, 2002, 296 (5567) : 550 - 553
[4] Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems
Caplen, NJ
Parrish, S
Imani, F
Fire, A
Morgan, RA
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (17) : 9742 - 9747
[5] Dopamine biosynthesis is selectively abolished in substantia nigra ventral tegmental area but not in hypothalamic neurons in mice with targeted disruption of the Nurr1 gene
Castillo, SO
Baffi, JS
Palkovits, M
Goldstein, DS
Kopin, IJ
Witta, J
Magnuson, MA
Nikodem, VM
[J]. MOLECULAR AND CELLULAR NEUROSCIENCE, 1998, 11 (1-2) : 36 - 46
[6] Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate
Elbashir, SM
Martinez, J
Patkaniowska, A
Lendeckel, W
Tuschl, T
[J]. EMBO JOURNAL, 2001, 20 (23) : 6877 - 6888
[7] Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
Fire, A
Xu, SQ
Montgomery, MK
Kostas, SA
Driver, SE
Mello, CC
[J]. NATURE, 1998, 391 (6669) : 806 - 811
[8] Genes and mechanisms related to RNA interference regulate expression of the small temporal RNAs that control C-elegans developmental timing
Grishok, A
Pasquinelli, AE
Conte, D
Li, N
Parrish, S
Ha, I
Baillie, DL
Fire, A
Ruvkun, G
Mello, CC
[J]. CELL, 2001, 106 (01) : 23 - 34
[9] A species of small antisense RNA in posttranscriptional gene silencing in plants
Hamilton, AJ
Baulcombe, DC
[J]. SCIENCE, 1999, 286 (5441) : 950 - 952
[10] Argonaute2, a link between genetic and biochemical analyses of RNAi
Hammond, SM
Boettcher, S
Caudy, AA
Kobayashi, R
Hannon, GJ
[J]. SCIENCE, 2001, 293 (5532) : 1146 - 1150

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