Selecting effective antisense reagents on combinatorial oligonucleotide arrays.
Nat Biotechnol
; 15(6): 537-41, 1997 Jun.
Article
in En
| MEDLINE
| ID: mdl-9181575
ABSTRACT
An array of 1,938 oligodeoxynucleotides (ONs) ranging in length from monomers to 17-mers was fabricated on the surface of a glass plate and used to measure the potential of oligonucleotide for heteroduplex formation with rabbit beta-globin mRNA. The oligonucleotides were complementary to the first 122 bases of mRNA comprising the 5' UTR and bases 1 to 69 of the first exon. Surprisingly few oligonucleotides gave significant heteroduplex yield. Antisense activity, measured in a RNase H assay and by in vitro translation, correlated well with yield of heteroduplex on the array. These results help to explain the variable success that is commonly experienced in the choice of antisense oligonucleotides. For the optimal ON, the concentration required to inhibit translation by 50% was found to be five times less than for any other ON. We find no obvious features in the mRNA sequence or the predicted secondary structure that can explain the variation in heteroduplex yield. However, the arrays provide a simple empirical method of selecting effective antisense oligonucleotides for any RNA target of known sequence.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Oligodeoxyribonucleotides
/
Globins
/
RNA, Messenger
Type of study:
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
Nat Biotechnol
Journal subject:
BIOTECNOLOGIA
Year:
1997
Document type:
Article
Affiliation country:
United kingdom