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In Vitro Method for Concerted Integration of Donor DNA Molecules Into Target DNA Using Retroviral Integrase and a Procedure for Evaluation of Donor DNA Integration.
Inventor: Dr. Duane P. Grandgenett
This technology relates to a method and a kit useful for the efficient integration of DNA donor molecules into DNA target molecules using retrovirus integrase (IN). Purified IN or integrase in virus particles and specifically designed donor DNA substrate having at least one unique restriction site for the analysis of concerted integration product, are used to mimic the integration of retrovirus A in vivo. For the production of transgenic animals and for gene transfer into cells, the donor DNA molecule is first complexed with IN prior to transfer into the target animals or cells. The formation of IN/donor DNA pre-integration complexes is the most efficient method to produce concerted integration recombinants with target DNA. In transgenics and gene transfer the host genome acts as the target DNA.
This technology also relates to an in vitro method for analyzing the concerted integration of DNA donor molecules into target DNA and the integrase enzymes that perform this integration reaction. Following the integration reaction, the reaction products are analyzed for at least one unique restriction site or genetic marker on the donor DNA molecule.
Integrases (IN) are important targets useful for the development and screening of anti-HIV therapeutic compounds. The method for concerted integration of DNA donor molecules into DNA target molecules using retroviral integrase together with the method for the detection of retrovirus integration into host DNA are useful for (i) ) production of transgenic animals, (ii) gene transfer in cultured cells, and, (iii) integrase screening of inhibitors targeting HIV-1 or HIV-2.
Keywords: Retrovirus, integrase, HIV
US Patent No. 5,811,270 (Date: Sept. 22, 1998);
US Patent 6,316,261 (Date: Nov. 13, 2001)
License: Collaborative research and licensing opportunities are available.
Reference Number: SLU-1006