Ctrc Research Foundation

The University of Texas System

Other

Ctrc Research Foundation Board of Regents

Bradford E Windle

Shih-Fong Chen

Terace M Fletcher

Ira Maine

Sean Michael Kerwin

Makoto Wajima

Blain Mamiya

Miquel Salazar

Miguel Salazar

Ming Qiu

The present invention relates to preparation and methods of use of novel extended and super-extended forms of DNA. These forms of DNA are stretched to a substantially linear form up to about 0.6 &mgr;m per kilobase pair. The extended DNA may be hybridized with suitably labelled probes and visualized directly. Gene order and distance are rapidly and efficiently determined with a resolution down to 0.4 kb using fluorescent hybridization techniques.

Methods and compositions for extended and super-extended DNA and hybridization mapping

It was found that normal human stem cells produce a regulated non-processive telomerase activity, while cancer cells produce a processive telomerase activity. Nucleotide analogs, such as 7-deaza-2&prime;-deoxyquanosine-5&prime;-triphosphate (7-deaza-dGTP) were found to be substrates for processive telomerase and incorporated into telomeric sequence. The incorporation of this nucleotide subsequently affected the processivity of telomerase, converting processive telomerase to non-processive telomerase. The incorporation of this nucleotide analogs was also found to inhibit formation of G-quartets by telomeric sequence. Other methods for converting cancer processive telomerase to the more benign non-processive telomerase include partially cleaving the telomerase RNA. The nucleoside analogs were found to be capable of a variety of activities including mediating allosteric-like inhibition of telomerase, premature termination and shortening of telomeric DNA, destabilization of telomeric structure and function and eventually cell death. Understanding the mechanisms of telomerase modulation by the 7-deaza-nucleotides has allowed the design of new telomerase inhibitors, modulators and agents for affecting telomere structure and function. These discoveries have application in the treatment of cancer.

Methods for modulation and inhibition of telomerase

It was found that normal human stem cells produce a regulated non-processive telomerase activity, while cancer cells produce a processive telomerase activity. Nucleotide analogs, such as 7-deaza-2'-deoxyquanosine-5'-triphosphate (7-deaza-dGTP) were found to be substrates for processive telomerase and incorporated into telomeric sequence. The incorporation of this nucleotide subsequently affected the processivity of telomerase, converting processive telomerase to non-processive telomerase. The incorporation of this nucleotide analogs was also found to inhibit formation of G-quartets by telomeric sequence. Other methods for converting cancer processive telomerase to the more benign non-processive telomerase include partially cleaving the telomerase RNA. The nucleoside analogs were found to be capable of a variety of activities including mediating allosteric-like inhibition of telomerase, premature termination and shortening of telomeric DNA, destabilization of telomeric structure and function and eventually cell death. Understanding the mechanisms of telomerase modulation by the 7-deaza-nucleotides has allowed the design of new telomerase inhibitors, modulators and agents for affecting telomere structure and function. These discoveries have application in the treatment of cancer.

Methods and compositions for modulation and inhibition of telomerase in

It was found that normal human stem cells produce a regulated non-processive telomerase activity, while cancer cells produce a processive telomerase activity. Nucleotide analogs, such as 7-deaza-2'-deoxyquanosine-5'-triphosphate (7-deaza-dGTP) were found to be substrates for processive telomerase and incorporated into telomeric sequence. The incorporation of this nucleotide subsequently affected the processivity of telomerase, converting processive telomerase to non-processive telomerase. The incorporation of this nucleotide analogs was also found to inhibit formation of G-quartets by telomeric sequence. Other methods for converting cancer processive telomerase to the more benign non-processive telomerase include partially cleaving the telomerase RNA. The nucleoside analogs were found to be capable of a variety of activities including mediating allosteric-like inhibition of telomerase, premature termination and shortening of telomeric DNA, destabilization of telomeric structure and function and eventually cell death. Understanding the mechanisms of telomerase modulation by the 7-deazanucleotides has allowed the design of new telomerase inhibitors, modulators and agents for affecting telomere structure and function. These discoveries have application in the treatment of cancer.

Improved telomerase activity assays are provided in which a ligation sequential reaction (LSR) or BrdUTP are used to identify a telomerase specific product. These assays are useful in diagnosing various cancers and determining the clinical prospects for cancer patients. In addition, the assays can be used to screen for substances that interfere with telomerase activity.

Rapid and sensitive assays for detecting and distinguishing between

The present invention relates generally to the fields of macromolecule image analysis and interpretation. More particularly, it concerns means for selecting an image, and using unique color vector computer automation to determine the shape, length, and physical characteristics of a stained DNA image midline based on the overall contour. The invention also includes methods of gravitationally stretching DNA to an essentially linear, 2-dimensional form having an inter kilobase pair distance of between 0.34 .mu.m to 0.65 .mu.m per kilobase pair. Examples of color images analyzed are presented and include the mapping of DIRVISH stained DNA markers, orientation, and distances. A novel use of the method allows determination of replication origin and termination sites on the DNA.

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San Antonio, TX, United States of America

Bradford E Windle