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Abstract

Disclosed are methods for suppressing or altering gene expression by locked nucleic acids that have high binding affinity for target sequences in genomic DNA. The present methods include a step of recombination to insert a foreign DNA fragment into a specific target site in a genome with greater efficiency than with current techniques. Using a protein to anchor a locked nucleic acid targeting construct at a specific site, DNA recombination between that site and foreign DNA is induced by employing the cell's innate repair machinery. As a result of this recombination, gene transcription can be effectively suppressed. The present methods work in all mammalian cells, and provide a rapid methodology to examine gene function. The foreign DNA can carry markers for ease of screening. The technique is applied in the production of stable cell lines with inserted or deleted genes.

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First Claim