Document Detail


Characterization of a potent and specific class of antisense oligonucleotide inhibitor of human protein kinase C-alpha expression.
MedLine Citation:
PMID:  9880552     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The use of antisense oligonucleotides to inhibit the expression of targeted mRNA sequences is becoming increasingly commonplace. Although effective, the most widely used oligonucleotide modification (phosphorothioate) has some limitations. In previous studies we have described a 20-mer phosphorothioate oligodeoxynucleotide inhibitor of human protein kinase C-alpha expression. In an effort to identify improved antisense inhibitors of protein kinase C expression, a series of 2' modifications have been incorporated into the protein kinase C-alpha targeting oligonucleotide, and the effects on oligonucleotide biophysical characteristics and pharmacology evaluated. The incorporation of 2'-O-(2-methoxy)ethyl chemistry resulted in a number of significant improvements in oligonucleotide characteristics. These include an increase in hybridization affinity toward a complementary RNA (1.5 degrees C per modification) and an increase in resistance toward both 3'-exonuclease and intracellular nucleases. These improvements result in a substantial increase in oligonucleotide potency (>20-fold after 72 h). The most active compound identified was used to examine the role played by protein kinase C-alpha in mediating the phorbol ester-induced changes in c-fos, c-jun, and junB expression in A549 lung epithelial cells. Depletion of protein kinase C-alpha protein expression by this oligonucleotide lead to a reduction in c-jun expression but not c-fos or junB. These results demonstrate that 2'-O-(2-methoxy)ethyl-modified antisense oligonucleotides are 1) effective inhibitors of protein kinase C-alpha expression, and 2) represent a class of antisense oligonucleotide which are much more effective inhibitors of gene expression than the widely used phosphorothioate antisense oligodeoxynucleotides.
Authors:
R A McKay; L J Miraglia; L L Cummins; S R Owens; H Sasmor; N M Dean
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  274     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1999 Jan 
Date Detail:
Created Date:  1999-02-11     Completed Date:  1999-02-11     Revised Date:  2012-06-25    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1715-22     Citation Subset:  IM    
Affiliation:
Department of Molecular Pharmacology, ISIS Pharmaceuticals, Carlsbad, California 92008, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Base Sequence
Dose-Response Relationship, Drug
Enzyme Inhibitors / administration & dosage,  pharmacology*
Gene Expression Regulation, Enzymologic / drug effects*
Humans
Isoenzymes / antagonists & inhibitors,  genetics*
Molecular Sequence Data
Oligodeoxyribonucleotides, Antisense / administration & dosage,  pharmacology*
Protein Kinase C / antagonists & inhibitors,  genetics*
Protein Kinase C-alpha
Proto-Oncogene Proteins c-fos / genetics,  metabolism
Proto-Oncogene Proteins c-jun / genetics,  metabolism
RNA, Messenger / metabolism
Tetradecanoylphorbol Acetate / pharmacology
Thermodynamics
Thionucleotides / administration & dosage,  pharmacology*
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Enzyme Inhibitors; 0/Isoenzymes; 0/Oligodeoxyribonucleotides, Antisense; 0/Proto-Oncogene Proteins c-fos; 0/Proto-Oncogene Proteins c-jun; 0/RNA, Messenger; 0/Thionucleotides; 151879-73-1/ISIS 3521; 16561-29-8/Tetradecanoylphorbol Acetate; EC 2.7.11.13/PRKCA protein, human; EC 2.7.11.13/Protein Kinase C; EC 2.7.11.13/Protein Kinase C-alpha

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  The expression of poly(A)-binding protein gene is translationally regulated in a growth-dependent fa...
Next Document:  Signal transducer and activator of transcription-1 and heat shock factor-1 interact and activate the...