DESCRIPTION:

Means for testing DNA cleavage presently require incubation with the enzyme followed by agarose or polyacrylamide gel electrophoresis to visually monitor changes in the DNA.  This usually takes 1 to 3 hours and requires a person to prepare, load and analyze the changes on the gel.  We are developing a new approach to monitoring DNA cleavage that would be fast and sensitive and could be automated for monitoring this process in many samples.  Enzymes that cleave DNA are found in many microorganisms including those that are pathogenic.  Thus, rapid procedures for monitoring specific DNA cleavage could be part of a testing protocol for samples potentially contaminated with pathogenic microorganisms. 

The method for monitoring DNA cleavage may also be useful for monitoring DNA binding.  DNA binding proteins regulate gene expression and when altered often lead to cancer.  This assay could potentially detect different levels of DNA binding proteins in tissue samples and be part of a diagnostic test for precancerous tissue samples.  The material we are using for detecting DNA cleavage and DNA binding are non-toxic to living cells, thus potentially allowing these assays to be used in monitoring these reactions in live organisms.

ADVANTAGES:

Present methodology for detecting cleavage of DNA is laborious and slow and can not be automated. This new method for detecting DNA cleavage allows for very fast sensitive and accurate measurements of the cleavage of DNA. DNA binding assays often take a minimum of 2-hours to complete. The developed assay could potentially be completed in a few minutes. In addition, gold nanoparticles are bio-compatible so that this method may also be viable for measuring DNA cleavage and DNA binding inside living cells.

DISADVANTAGES:

The group has not yet tested all of the conditions possible that are compatible with the Aunanoparticles and the action of the restriction enzymes and DNA binding proteins.

POTENTIAL APPLICATIONS:

Companies that are potentially interested in this technology include those from computer chip, microelectronics, pharmaceuticals, chemical sensor/biosensor industries, and national security applications.

PATENT STATUS:

Patent strategy is being evaluated