Abstract: The present invention is directed to novel telomerase nucleic acids and amino acids. In particular, the present invention is directed to nucleic acid and amino acid sequences encoding various telomerase protein subunits and motifs, including the 123 kDa and 43 kDa telomerase protein subunits of Euplotes aediculatus, and related sequences from Schizosaccharomyces, Saccharomyces sequences, and human telomerase. The present invention is also directed to polypeptides comprising these telomerase protein subunits, as well as functional polypeptides and ribonucleoproteins that contain these subunits.

Abstract: Methods and materials for culturing primate-derived primordial stem cells are described. In one embodiment, a cell culture medium for growing primate-derived primordial stem cells in a substantially undifferentiated state is provided which includes a low osmotic pressure, low endotoxin basic medium that is effective to support the growth of primate-derived primordial stem cells. The basic medium is combined with a nutrient serum effective to support the growth of primate-derived primordial stem cells and a substrate selected from the group consisting of feeder cells and an extracellular matrix component derived from feeder cells. The medium further includes non-essential amino acids, an anti-oxidant, and a first growth factor selected from the group consisting of nucleosides and a pyruvate salt.

Abstract: This invention provides purified telomerase and methods of purifying it. The methods involve the use of several sequential steps, including the use of matrices that bind molecules bearing negative charges, matrices that bind molecules bearing positive charges, intermediate-selectivity matrices, methods that separate molecules based on their size, shape, or buoyant density, and by affinity purification.

Abstract: The present invention is related to novel nucleic acids comprising telomerase reverse transcriptase (TERT) cis-acting transcriptional control sequences, including TERT human and mouse promoter sequences. The present invention is further directed to methods of using these cis-acting transcriptional control sequences, for example, to drive heterologous gene sequences; to modulate the level of transcription of TERT or to isolate novel trans-acting regulatory factors which bind to and modulate the activity of a TERT promoter.

Abstract: This invention provides for murine telomerase reverse transcriptase (mTERT) enzyme proteins and nucleic acids, including methods for isolating and expressing these nucleic acids and proteins, which have application to the control of cell proliferation and aging, including the control of age-related diseases, such as cancer.

Abstract: The present invention is related to novel nucleic acids comprising telomerase reverse transcriptase (TERT) cis-acting transcriptional control sequences, including TERT human and mouse promoter sequences. The present invention is further directed to methods of using these cis-acting transcriptional control sequences, for example, to drive heterologous gene sequences; to modulate the level of transcription of TERT or to isolate novel trans-acting regulatory factors which bind to and modulate the activity of a TERT promoter.

Abstract: This disclosure provides a system for specifically killing cancer cells which can be used in the course of human therapy. Vectors of the invention comprise an encoding sequence for a glycosyltransferase, under control of a tumor or tissue specific transcriptional control element, such as the promoter for telomerase reverse transcriptase. Exemplary glycosyltransferases are the A or B transferase enzymes, which cause the cancer cells to express ABO histo blood group allotypes against which humans have naturally occurring antibody. This provides for ongoing surveillance for newly emerging cells with a malignant phenotype.

Abstract: Pregnancy in animals such as the pig usually does not continue to term when there are only a few embryos in the uterus. Currently available technology for maintaining pregnancy involves a series of hormone injections, and the failure rate is high. It has now been discovered that pregnancy can be maintained by complementing fertile embryos in the uterus with additional embryos that aren't expected to survive the full term of the pregnancy. Particularly suitable are parthenogenetic embryos, formed by activating and diploidizing unfertilized oocytes. The carrier embryos are engrafted into the uterus of a surrogate female, and the pregnancy continues to term without further intervention—even if the number of fertile embryos are below the minimum litter size. This provides valuable biological material that can be used for transplantation, the production of pharmaceuticals, and for agricultural use.

Abstract: This disclosure provides a system for obtaining expression libraries from primate pluripotent stem (pPS) cells. pPS cells can be maintained in vitro without requiring a layer of feeder cells to inhibit differentiation. The role of the feeder cells is replaced by several other culture conditions provided in a suitable combination. Conditions that promote pPS cell growth without differentiation include supporting the culture on an extracellular matrix, and culturing the cells in a medium conditioned by another cell type. The cDNA libraries from such cultures are devoid of transcripts of feeder cell origin, relatively uncontaminated by transcripts from differentiated cells, and can have a high proportion of full-length transcripts. Subtraction libraries can also be produced that are enriched for transcripts modulated during differentiation.

Abstract: Oligonucleotides with a novel sugar-phosphate backbone containing at least one internucleoside 3′-NHP(O)(S−)O-5′ linkage, and methods of synthesizing and using the inventive oligonucleotides are provided. The inventive thiophosphoramidate oligonucleotides were found to retain the high RNA binding affinity of the parent oligonucleotide N3′→P5′ phosphoramidates and to exhibit a much higher acid stability.

Abstract: This invention provides media that support the growth of primate pluripotent stem cells in feeder-free culture, and cell lines useful for producing such media and other purposes. Conventionally, it has been necessary to grow pluripotent embryonic cells on feeder layers of primary embryonic fibroblasts, in order to prevent them from differentiating. It has now been discovered that standard culture media conditioned by special cell lines can be used to support proliferation of pluripotent stem cells while inhibiting differentiation in an environment free of feeder cells. This invention includes mesenchymal and fibroblast-like cell lines obtained from embryonic tissue or differentiated from embryonic stem cells. Methods for deriving such cell lines, processing media, and growing stem cells using the conditioned media are described and illustrated in this disclosure.

Abstract: The present invention provides TRT antisense oligonucleotides, methods of detecting TRT, methods of diagnosing telomerase-related conditions, methods of diagnosing and providing a prognosis for cancer, and methods of treating telomerase-related conditions, including cancer.

Abstract: The invention provides compositions and methods related to human telomerase reverse transcriptase (hTRT), the catalytic protein subunit of human telomerase. The polynucleotides and polypeptides of the invention are useful for diagnosis, prognosis and treatment of human diseases, for changing the proliferative capacity of cells and organisms, and for identification and screening of compounds and treatments useful for treatment of diseases such as cancers.

Abstract: The invention provides compositions and methods related to human telomerase reverse transcriptase (hTRT), the catalytic protein subunit of human telomerase. The polynucleotides and polypeptides of the invention are useful for diagnosis, prognosis and treatment of humana diseases, for changing the proliferative capacity of cells and organisms, and for identification and screening of compounds and treatments useful for treatment of diseases such as cancers.

Abstract: Oligonucleotides with a novel sugar-phosphate backbone containing at least one internucleoside 3′-NHP(O)(S−)O-5′ linkage, and methods of synthesizing and using the inventive oligonucleotides are provided. The inventive thiophosphoramidate oligonucleotides were found to retain the high RNA binding affinity of the parent oligonucleotide N3′→P5′ phosphoramidates and to exhibit a much higher acid stability.

Abstract: A new protein named Tankyrase II is described in this disclosure. Sequences for the human Tankyrase II cDNA and the protein translation product are provided. Also provided are species homologs, muteins, related nucleic acids, peptides, and drug screening assays. Tankyrase II interacts with telomere-associated proteins, thereby affecting telomerase activity and potentially telomere length. The materials and techniques provided in this disclosure allow Tankyrase II activity to be studied in vitro and manipulated inside cells—to the potential benefit of clinical conditions associated with a defect in telomerase activity, or the replicative capacity of affected cells.

Abstract: This invention provides a system for producing differentiated cells from a stem cell population for use wherever a relatively homogenous cell population is desirable. The cells contain an effector gene under control of a transcriptional control element (such as the TERT promoter) that causes the gene to be expressed in relatively undifferentiated cells in the population. Expression of the effector gene results in depletion of undifferentiated cells, or expression of a marker that can be used to remove them later. Suitable effector sequences encode a toxin, a protein that induces apoptosis, a cell-surface antigen, or an enzyme (such as thymidine kinase) that converts a prodrug into a substance that is lethal to the cell. The differentiated cell populations produced according to this disclosure are suitable for use in tissue regeneration, and non-therapeutic applications such as drug screening.

Abstract: Method and compositions are provided for the determination of telomere length and telomerase activity, as well as the ability to increase or decrease telomerase activity in the treatment of proliferative diseases. Particularly, primers are elongated under conditions which minimize interference from other genomic sequences, so as to obtain accurate determinations of telomeric length or telomerase activity. In addition, compositions are provided for intracellular inhibition of telomerase activity and means are shown for slowing or reversing the loss of telomeric repeats in aging cells.

Abstract: Nucleic acids comprising the RNA component of a mammalian telomerase are useful as pharmaceutical, therapeutic, and diagnostic reagents.

Abstract: An optimum time has been discovered for obtaining porcine oocytes with improved developmental competence. Harvested oocytes are matured by culturing in vitro, and then activated ˜42-46 hours after beginning of the culture period. Alternatively, ovulation is induced in a donor female using a gonadotrophic hormone (optionally monitored by ultrasonography), maturation is allowed to proceed in vivo, and then the harvested oocyte is activated ˜44-48 hours after inducing ovulation. Outside the optimal time frame, the ability of the oocytes to undergo parthenogenetic activation declines markedly. These methods overcome problems encountered with previous protocols for nuclear transfer and assisted reproduction in the pig.