Targeted mutagenesis in living cells using modified oligonucleot

Details Targeted mutagenesis in living cells using modified oligonucleot Targeted mutagenesis in living cells using modified oligonucleot

1997-03-26

2000-10-24

LeGuyander, John L.

Chemistry: molecular biology and microbiology

Animal cell, per se ; composition thereof; process of...

Method of regulating cell metabolism or physiology

435 6, 536 243, 536 2431, 536 2433, 536 245, C12Q 168, C07H 2104

Patent

active

061366013

ABSTRACT:
A method for introducing a site-specific mutation into a target polynucleotide sequence is presented. The method involves the use of an oligonucleotide capable of binding to the target sequence, either by triplex formation (mediated by Hoogsteen, reverse Hoogsteen or equivalent base pairing) or by Watson/Crick base pairing (in the presence of a recombinase enzyme). The oligonucleotide of the invention is modified by the covalent attachment of one or more electrophilic groups. When a modified oligonucleotide is bound to its target sequence, the electrophilic group is able to interact with a nearby nucleotide in the target sequence, causing a modification to the nucleotide that results in a change in nucleotide sequence. Compositions used in the practice of the method are also disclosed.

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