Development of a Single Vector Containing cre Recombinase and Two Functional lox Sites: Active cre Produced in Eukaryotic but not Prokaryotic Systems
Description of Invention:
The bacterial recombinase cre will recombine lox sites within bacteria. Since this will occur with extremely low levels of cre, it has not been possible to place the cre gene within a vector which also contains two lox recombination site and clone the construct in bacteria. Therefore, in order to use cre-lox technology, the use of two separate vectors has been required -- one containing cre and another containing the lox sites. The inventors have devised a strategy to generate vectors that contain cre in a form which is not translated in bacteria thereby allowing for the co-existence of two lox sites within the same vector. Under these circumstances, the vector can be cloned and grown in bacteria enabling experiments to be conducted in eukaryotic cells using just one vector instead of two separate vectors. This system provides a significant advantage in performing many types of experiments.
In in vitro transfection experiments, only one vector needs to be incorporated into a cell instead of two separate vectors. This overcomes the inherent problem of trying to transfect two vectors into one cell, where the relative ratios of the two vectors which enter the cells can vary widely. Of even greater significance is the application of this technology to transgenic animal work where the incorporation of one vector into a line of transgenic animals is all that is required, instead of the generation of two separate lines of transgenic animals which then must be crossed to produce an animal which contains both constructs. In addition, this technology can be applied to gene therapy approaches in which the tissue-specific expression of a therapeutic gene can be activated by cre contained within the same construct. The technology allows generation of one vector which contains the cre-variant and two lox sites enabling one to either switch the expression of one gene to another gene and/or amplify the expression of a particular gene to high levels in a tissue specific manner.
Inventors:
Stan Kaczmarczyk (NCI) Jeffrey E. Green (NCI)
Patent Status:
DHHS Reference No. E-172-2000/0 -- Research Materials
Relevant Publication: SJ Kaczmarczyk and JE Green. A single vector containing modified cre recombinase and LOX recombination sequences for inducible tissue-specific amplification of gene expression. Nucleic Acids Res. 2001 Jun 15;29(12):E56-E66. [PubMed abs]
Licensing Status: Available for non-exclusive licensing under a Biological Materials License Agreement.
Portfolios: Gene Based Therapies
Gene Based Therapies -Therapeutics-Gene Therapy-Vectors Gene Based Therapies -Therapeutics
For Additional Information Please Contact: Jennifer Wong
NIH Office of Technology Transfer
6011 Executive Blvd, Suite 325
Rockville, MD 20852-3804
Phone: (301)435-4633
Email: wongje@mail.nih.gov
Fax: (301)402-0220
