Columbia University

Technology Ventures

Method for library construction by generating directed, recombinant fusion nucleic acids

Technology #646

“Lead Inventor: Rodney Rothstein Ph.D.

Library for genetic recombination and genomic sequences:
The use of libraries is widespread in research and in the pharmaceutical industry. In general, library construction involves many steps before the production of a final, usable library. Conventional approaches for generating recombinant sequence are limited by low integration and recombination frequencies. This invention developed a method of library construction that relies on genetic recombination and the availability of complete or partial genomic sequences, which provides a novel and more efficient method of library construction.

Genomic sequence libraries:
The present invention provides for a method for generating a directed, recombinant nucleic acid molecule. This invention also provides methods for producing fusion nucleic acid molecule capable of crossover recombination. This comprises (a) using two pairs of primers to amplify, separately, two double-stranded nucleic acid molecules by PCR; (b) combining amplification products with first primer of the first pair and second primer of second pair; and © amplifying the hybridized products to generate directed, recombinant fusion nucleic acid. This invention also provides for the use of adaptamers to design or manipulate fragments for recombination-directed library construction.

Applications:
• Construct libraries for studying gene function
• Direct replacement of particular genes, which might aid future gene therapy development
• Two-hybrid library could be built to analyze linkage between genes and crosstalk of different pathways to potentially generate new leads for drug discovery
• Specific protein tags (GFP) could be engineered to fuse with all the proteins in an organism to screen for compounds that affect a series of genes

Advantages:
• Production of libraries can now be performed in a more efficient manner without use of intermediate host and without requiring ligation
• It allows a complete arrayed library to be formed rather than specific gene constructs
• The adaptamers are designed to be specific for just one other adaptamer and will not amplify any internal sequences, so new sequences can be fused to any other selected fragment


Patent Status: Patents Issued (US 5,942,422; US 6,291,213) ~ see links below.

Licensing Status: Available for Licensing and Sponsored Research Support