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Nucleic Acids Research, 1994, Vol. 22, No. 12 2366-2369
© 1994

CHEMISTRY

Enhancement of selectivity in recognition of nucleic acids via chemical autoligation

Sergei M. Gryaznov*,, Ronald Schultz1, Surendra K. Chaturvedi1 and Robert L. Letsinger1

Lynx Therapeutics Inc. 3832 Bay Center Place, Hayward, CA 94545 1Department of Chemistry, Northwestern University Evanston, IL 60208, USA

*To whom correspondence should be addressed

Received February 17, 1994. Revised May 2, 1994. Accepted May 2, 1994.

A new approach to increase the selectivity of interaction between oligonucleotide probes and target nucleic acids is described. In place of a single, relatively long oligonucleotide probe, two or three short oligomers terminated by thiophosphoryl and bromoacetamido groups are employed. Fast and efficient autoligation takes place when the oligomers hybridize in a contiguous mode to the same complementary strand such that a thiophosphoryl group on one strand and a bromoacetamido group on another are brought into proximity. A single nucleotide mismatch for the short probes leads to marked reduction in the rate of autoligation. The binding affinity of the product is close to that for a natural probe of the same length. This approach could have potential in oligonucleotide-based diagnostics, chemical amplification systems, and therapeutic applications.


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