Complex host cell responses to antisense suppression of ACHE gene expression

Citation
N. Galyam et al., Complex host cell responses to antisense suppression of ACHE gene expression, ANTISENSE N, 11(1), 2001, pp. 51-57
Citations number
30
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Molecular Biology & Genetics
Journal title
ANTISENSE & NUCLEIC ACID DRUG DEVELOPMENT
ISSN journal
1087-2906 → ACNP
Volume
11
Issue
1
Year of publication
2001
Pages
51 - 57
Database
ISI
SICI code
1087-2906(200102)11:1<51:CHCRTA>2.0.ZU;2-V
Abstract
3'-End-capped, 20-mer antisense oligodeoxynucleotides (AS-ODN) protected wi th 2'-O-methyl (Me) or phosphorothioate (PS) substitutions were targeted to acetylcholinesterase (AChE) mRNA and studied in PC12 cells. Me-modified AS -ODN suppressed AChE activity up to 50% at concentrations of 0.02-100 nM PS -ODN was effective at 1-100 nM. Both AS-ODN displayed progressively decreas ed efficacy above 10 nM, In situ hybridization and confocal microscopy demo nstrated dose-dependent decreases, then increases, in AChE mRNA, Moreover, labeling at nuclear foci suggested facilitated transcription or stabilizati on of AChE mRNA or both under AS-ODN, Intracellular concentrations of bioti nylated oligonucleotide equaled those of target mRNA at extracellular conce ntrations of 0.02 nM yet increased only 6-fold at 1 muM ODN. Above 50 nM, s equence-independent swelling of cellular, but not nuclear, volume was obser ved. Our findings demonstrate suppressed AChE expression using extremely lo w concentrations of AS-ODN and attribute reduced efficacy at higher concent rations to complex host cell feedback responses.