I ran across Health Reseach Inc.'s Technology Transfer website and thought that maybe somone would be interested in some of these opportunities. For the full list go to:

http://www.hrinet.org/



Technology Opportunity
Invention Available for Licensing and Commercialization



Universal Reverse Transcription and PCR Designed for Human Tissue mRNA-Specific Amplification and Quantitation




Description

A new method has been developed to detect gene expression at the mRNA level in extracts from tissue specimens, without confounding by contaminating genomic DNA encoded pseudogenes. The technology makes true quantitation of gene expression in human tissue samples of any size accurate and feasible.



Researchers have devised a novel reverse transcription strategy that utilizes an mRNA-specific RT primer designed to be universal for all mRNA transcripts. The employment of the mRNA-specific RT strategy obviates the limitations posed by total RNA preps contaminated with gDNA. Subsequent PCR is performed integrating features of the universal RT primer design, and is not complicated by the presence of gDNA nor pseudogene. Message RNA can thus be amplified very specifically in the setting of gDNA contamination, without further steps. Purification of total RNA to mRNA becomes unnecessary, a particularly critical advantage when handling small tissue samples, as in human tissues. Both DNase treatment after RNA isolation, and no-RT controls in the PCR step to assess the presence of pseudogene, become unnecessary. Standard reference housekeeper transcripts such as β-actin and GAPDH, otherwise confounded by gDNA-encoding pseudogenes, can be amplified specifically and quantitatively. Multiple transcripts can be amplified from the same total RNA aliquot, maximizing the efficiency of reverse transcription on limited RNA samples.



The universal reverse transcription strategy is quantitatively similar in efficiency to conventional odT techniques. PCR primer design is not further complicated by the presence of pseudogene sequences competing for PCR primer. Genomic DNA encoded pseudogene sequences that exist for reference gene transcripts (e.g. β-actin and GAPDH) and target gene transcripts (e.g. GSTM1 and GSTP1) have been amplified alongside nonpseudogene-encoded sequences (e.g. CYP1B1, NQO1) by otherwise conventional PCR techniques, with the only change being the use of different RT and PCR primers.



The system has been readily employed in the common and currently used qualitative and real-time quantitative PCR protocols assessing gene expression in small samples of human tissues. The strategy has therefore proven to be universally applicable to all tested transcripts in all tested gene-expression assay contexts.

This diagnostic method monitors changes in viral tropism associated with HIV disease progression, and can be used as a diagnostic tool to monitor the effectiveness of antiviral therapy and disease progression in HIV positive individuals.



A change in viral tropism occurs in many HIV positive individuals over time, and this change is indicated by a shift in coreceptor usage that has been shown to correlate with increased disease pathogenesis. Therefore, application of the diagnostic method to monitor shifts in coreceptor usage is useful for predicting disease progression.



Understanding coreceptor usage during potent antiretroviral therapy is relevant to HIV-1 dynamics and the maintenance of viral suppression and clinical response. The method utilizes a mathematical model to measure the proportion of virus in a specimen that uses each coreceptor. Coreceptor utilization studies may have a role in virologic monitoring of patients on antiviral therapy, particularly as drugs are developed which target R5 or X4 strains of HIV-1. The diagnostic method and the mathematical model provide a laboratory technique to monitor the effectiveness of antiviral therapy.
Potential Market Applications

Gene expression Human gene expression profiling

Genetic regulation Quantitative RT-PCR

Expression Arrays Gene-environment interaction


Main Advantages of Invention

1. Universal reverse transcription for all tested transcripts.

2. Message-RNA-specific PCR.

3. Standard RNA isolation techniques are acceptable no additional purification steps.

4. Multiple transcripts can be amplified from one total RNA sample.

5. Standard housekeeper reference transcripts (e.g. β-actin, GAPDH) are valid controls.

6. Useable with standard RT-PCR strategies/kits only primers differ.

7. Quantitation of low copy transcripts in small tissue samples is feasible.

8. Sensitivity similar to standard RT-PCR protocols.




State of Development

1. Fully operational universal reverse transcription primer at research scale.

2. PCR primers specifically designed for >12 transcripts also at research scale.

3. Interindividual (n>50) sample performance verified, and subset quantified.

4. Performance versus standard odT strategy quantified.

5. All relevant controls demonstrated.



Potential Areas of Application

Gene expression assays, from traditional to microarrays.

Quantitative RT-PCR applications.

Gene regulation research.

Gene expression phenotyping of cells and tissues for research or clinical molecular pathology.

Small sample (e.g. microdissection) or single cell expression assays.

Interindividual expression phenotyping for disease susceptibility.

Licensing Potential

HRI seeks commercial partners to develop the technology for clinical and commercial use. Available for licensing.





Contact:

Bob Gallo

Associate Director Technology Transfer

Health Research, Inc.

One University Place

Rensselaer, NY 12144-3456

(518) 431-1208 Fax (518) 431-1234
rlg04@health.state.ny.us