75 results about "Telomerase reverse transcriptase" patented technology

The present invention is in the fields of molecular and cellular biology. The invention is generally related to reverse transcriptase enzymes and methods for the reverse transcription of nucleic acid molecules, especially messenger RNA molecules. Specifically, the invention relates to reverse transcriptase enzymes which have been mutated or modified to increase thermostability, decrease terminal deoxynucleotidyl transferase activity, and / or increase fidelity, and to methods of producing, amplifying or sequencing nucleic acid molecules (particularly cDNA molecules) using these reverse transcriptase enzymes or compositions. The invention also relates to nucleic acid molecules produced by these methods and to the use of such nucleic acid molecules to produce desired polypeptides. The invention also concerns kits comprising such enzymes or compositions.

This invention relates to pork liver cell immortalities and its producing method. Procedures are showed: high live ratio fresh original pork liver cell is got by dispase-collagenase perfusion method; it is cultured for 24 hours. Then cell upper heat removing of recombination retronituse that contains SV40 big T antigen is used to infect the pork cell under condition of polybrene which concentration is 8ug / ml. Then it is medicine pressure filtrated by 500ug / ml G418 after one week, single clone cell is picked for large culture when the cell clone appears and grows to 1.0-2.0cm to get pork cell that can passage. Reinfection is done under condition of 8ug / ml polybrene. Then it is medicine pressure filtrated by 2ug / ml puromycin after one week. Single clone cell is picked for large culture when the cell clone appears and grows to 1.0-2.0cm to get immortality pork cell.

Compounds, and methods of using the same, are provided which increase the expression of telomerase reverse transcriptase (TERT) in a cell. These compounds and methods find use in a variety of applications in which increased expression of telomerase is desired, including immortalization of cell lines and treating conditions in a subject characterized by cellular senescence.

The present invention provides a versatile mutant reverse transcriptase with high thermal stability, a nucleic acid thereof and a method for producing a mutant reverse transcriptase, a versatile kits for reverse transcription and detection, a method for improving thermal stability of a nucleic acid-related enzyme, which significantly improves thermal stability of a nucleic acid-related enzyme, and a reverse transcription method, which efficiently performs a reverse transcription. An amino acid residue in a nucleic acid interaction region of a wild-type enzyme is substituted with a positively-charged amino acid residue or a nonpolar amino acid residue, to form a nucleic acid interaction region having a positive effective charge larger than the nucleic acid interaction region of a wild-type enzyme.

Methods and compositions are provided for modulating, e.g., increasing or decreasing, the expression of telomerase reverse transcriptase (TERT). In the subject methods, the binding interaction of the TERT Site C repressor site with a Site C repressor protein complex made up of one or more proteins is modulated to achieve the desired change in TERT expression. A feature of the subject invention is that the target Site C repressor protein complex includes an LSF protein. The subject methods and compositions find use in a variety of different applications, including the immortalization of cells, the production of reagents for use in life science research, therapeutic applications; therapeutic agent screening applications; and the like.

The invention provides dendritic cell (DC) preparations that present a telomerase reverse transcriptase (TRT) peptide in the context of an MHC class I or MHC class II molecule. The DCs may be pulsed with a TRT polypeptide, or may comprise a recombinant polynucleotide encoding TRT. The invention also describes the use of such compositions for the prevention and treatment of cancers and other diseases.

The present invention relates to preadipocyte strains that maintain replicative potential and adipogenic capacity. In particular, the invention relates to preadipocytes engineered to express telomerase reverse transcriptase (TERT). Use of the cells as research tools, in screening assays, and as therapeutic and / or clinical reagents is also described.

The present invention is in the fields of molecular and cellular biology. The invention is generally related to reverse transcriptase enzymes and methods for the reverse transcription of nucleic acid molecules, especially messenger RNA molecules. Specifically, the invention relates to reverse transcriptase enzymes which have been mutated or modified to increase thermostability, decrease terminal deoxynucleotidyl transferase activity, and / or increase fidelity, and to methods of producing, amplifying or sequencing nucleic acid molecules (particularly cDNA molecules) using these reverse transcriptase enzymes or compositions. The invention also relates to nucleic acid molecules produced by these methods and to the use of such nucleic acid molecules to produce desired polypeptides. The invention also concerns kits comprising such enzymes or compositions.

An objective of the present invention is to provide an adenovirus vector expressing a REIC / Dkk-3 protein at a high level and containing a DNA construct for expression of REIC / Dkk-3 DNA, wherein the DNA construct is prepared by ligating, from the 5′ terminal side,(i) a CMV promoter,(ii) REIC / Dkk-3 DNA,(iii) a polyA addition sequence, and(iv) enhancers prepared by linking an hTERT (Telomerase Reverse Transcriptase) enhancer, an SV40 enhancer, and a CMV enhancer in this order.

The present invention is directed to pharmaceutical compositions comprising a telomerase reverse transcriptase polypeptide or a polypeptide homologous to a telomerase reverse transcriptase. The present invention is also directed to pharmaceutical compositions comprising a polynucleotide encoding either of the aforesaid polypeptides. The present invention is further directed to methods for eliciting an immune response to telomerase reverse transcriptase in a subject.

Polynucleotides encoding telomerase reverse transcriptase (TERT) fusion proteins are provided, the TERT fusion proteins comprising a TERT protein, or functional variant thereof, fused to a substantialportion of the B subunit of heat labile enterotoxin (LTB). TERT variants useful in TERT-LTB fusion proteins of the invention comprise mutations that function to eliminate telomerase catalytic activity. The polynucleotides of the present invention can elicit an immune response in a mammal, which, in preferred embodiments, is stronger than the immune response elicited by a wild-type TERT. TERT expression is commonly associated with the development of human carcinomas. The present invention provides compositions and methods to elicit or enhance immunity to the protein product expressed by the TERT tumor-associated antigen, wherein aberrant TERT expression is associated with a carcinoma or its development. This invention specifically provides adenoviral vector and plasmid constructs carryingpolynucleotides encoding TERT fusion proteins and TERT variants and discloses their use in vaccines and pharmaceutical compositions for preventing and treating cancer.

Stabilized reverse transcriptase fusion proteins including a thermostable reverse transcriptase connected to a stabilizer protein are described. Attaching the stabilizer protein to the thermostable reverse transcriptase stabilizes the fusion protein and can aid in its purification, provide increased solubility, allow for longer storage, or allow the fusion protein to be used under more rigorous conditions such as higher temperature. The stabilized reverse transcriptase fusion protein can also include a linker between the stabilizer protein and the thermostable reverse transcriptase. The stabilized reverse transcriptase fusion proteins are suitable for use in nucleic acid amplification methods such as the reverse transcription polymerase chain reaction and other applications involving cDNA synthesis.

The invention belongs to vaccine and preparation thereof, in particular to a vaccine for preventing and curing tumor and preparation thereof. The vaccine is prepared by steps of: choosing a telomerase reverse transcriptase epitope sequence, serially connecting nucleotides the telomerase reverse transcriptase with multi-epitope, preparing the DNA vaccine of the telomerase reverse transcriptase with multi-epitope, and preparing the multi-epitope vaccine of the telomerase reverse transcriptase with multi-epitope. The invention overcomes the shortcomings that a single peptide vaccine only has a clinical effect in several tumor patients. After the vaccine has an immune body, the epitope specific CD4+T cells and the CD8+T cells can be activated. Under the assistance of the Th1 cell factor excreted from the CD4+T cells, the CD8+CTL can kill the tumor cell presenting TERT so as to prevent and cure tumor.

The present invention relates to a reverse transcriptase having improved thermostability, more precisely a mutant reverse transcriptase with improved thermostability by substitution of one or more amino acids selected from the group consisting of the 63rd glutamine (Q63), the 264th lysine (K264), the 295th lysine (K295), the 306th threonine (T306), the 346th glutamic acid (E346), the 408th proline (P408), the 438th histidine (H438), and the 454th asparagin (N454) of the amino acid sequence of M-MLV originated reverse transcriptase represented by SEQ. ID. NO: 1 with other amino acids. The mutant reverse transcriptase of the present invention demonstrates excellent thermostability, compared with the wild type reverse transcriptase. Therefore, it is advantageous to obtain the target cDNA with stable reverse transcription activity even in the presence of RNA that can form the stable secondary structure at a high temperature.

The invention relates to an ORFV virulence weakening method. A calf testicle cell line is utilized to carry out continuous passage culture of the ORFV, so that the ORFV virulence is weakened, thereby providing a stable and reliable method for preparing and producing an ORFV virulence weakening vaccine; and the method is used for obtaining abundant stable vaccines. The method disclosed by the invention comprises the following steps: 1) separating and initially culturing the ORFV; and 2) carrying out continuous passage culture on the ORFV in the calf testicle cell line until the virulence is weakened. In the step 1), the initial culture of the ORFV adopts the calf testicle cell line, and the culture fluid of the calf testicle cell line is M199, DMEM (Dulbecco Modified Eagle Medium) (high-sugar) and RPMI-1640 culture media. In the step 1), the ORFV is separated from lip scab of a sicken goat. In the step 2), the calf testicle cell line is prepared by introducing a telomerase reverse transcriptase (hTERT).

A method for enhancing the in vitro and in vivo lifespan of T cells is provided. A method for enhancing the life span of a T cell is provided. The method includes engineering the T cell to express exogenous ribonucleic acid (RNA), where the RNA includes a nucleic acid encoding telomerase reverse transcriptase (TERT). T cells genetically engineered to transiently express exogenous RNA which includes the coding sequence for TERT (TERT T cells) are provided. TERT T cells have one or more of the following characteristics when compared to a T cells not expressing TERT from an exogenously introducednucleotide: enhanced in vitro and in vivo proliferation, decreased number of senescent cells, and enhanced in vivo anti-cancer activity. The modified TERT T cells can be used in adoptive cell transfer to treat a subject in need thereof.

Methods and compositions are provided for modulating, and generally upregulating, the expression of telomerase reverse transcriptase (TERT) by blocking repression of TERT transcription, e.g., by inhibiting binding of repressor factor to a Site C repressor binding site located in the TERT minimal promoter. The subject methods and compositions find use in a variety of different applications, including the immortalization of cells, the production of reagents for use in life science research, therapeutic applications; therapeutic agent screening applications; and the like. In further describing the subject invention, the methods and compositions of the invention are described first in greater detail, followed by a review of the various applications in which the subject invention finds use.

The invention discloses a kit for detecting TERT (telomerase reverse transcriptase) gene mutation, and a detection method of the kit. The kit comprises an LNA (locked nucleic acid) modified specific probe for a TERT gene mutation site, wherein the specific probe can be combined with wild DNA (deoxyribonucleic acid), and can detect a sample containing 0.01% TERT gene mutation DNA; and a minimum detection limit is 2-5cp. The detection method for detecting the TERT gene mutation has the advantages of high specificity, high sensitivity, low pollution, simplicity and rapidness in operation, safety and the like, is especially suitable for detecting the gene mutation from body fluids containing low mutation content such as plasma, urine and saliva, is suitable for performing early screening and diagnosis on a bladder cancer, and provides guidance for personalized medicine.

The invention discloses an application of Bst DNA polymerase in RNA amplification. The Bst DNA polymerase has reverse transcriptase activity, and thus RNA is subjected to inverse transcription into cDNA under action of the Bst DNA polymerase. A new RNA amplification detection method is established on the basis of the new discovery that the Bst DNA polymerase has reverse transcriptase activity. The method integrates inverse transcription and subsequent nucleic acid amplification, temperature adjustment is not needed during the whole process, addition of extra reverse transcriptases is not needed, operation is simple, reaction time is shortened, and new methodology reference and technical support are provided for RNA inverse transcription and amplification detection. The discovered Bst DNA polymerase can perform an inverse transcription reaction at a high temperature, which facilitates to open and combine RNAs with complex secondary structures, and therefore the problem is solved that reverse transcriptases of AMV or the like are difficult to amplify RNA advanced structure areas.

Methods and compositions are provided for modulating, and generally upregulating, the expression of telomerase reverse transcriptase (TERT) by blocking repression of TERT transcription, e.g., by inhibiting binding of repressor factor to a Site C repressor binding site located in the TERT minimal promoter. The subject methods and compositions find use in a variety of different applications, including the immortalization of cells, the production of reagents for use in life science research, therapeutic applications; therapeutic agent screening applications; and the like. In further describing the subject invention, the methods and compositions of the invention are described first in greater detail, followed by a review of the various applications in which the subject invention finds use.

The invention provides a building method of a transposable element carrier expressing pig telomerase reverse transcriptase, and application in building a pig immortalization cell line, and belongs to the field of biotechnology research. The building method includes the steps of cloning of a promoter of a pig protein translation elongation factor 1a and detection of transcriptional activity, building of an SB transposable element expression vector and exogenous gene integration, pig telomerase reverse transcriptase cDNA cloning, activity detection and building of the pig fibroblast immortalization cell line. Compared with other methods, the pig immortalization cell line built through the transposable element carrier comprises non-transformed normal cells, does not contain resistance genes, virogenes or oncogenes, and is suitable for cell physiological function researching, pig virus separation cultivation and vaccine production.

The invention relates to a method for building the immortalized preadipocytes of chicken, which can be used for solving the problems that the stable and reliable result is difficult to obtain by the research as the preadipocytes of primary chicken can not go down to the future generation infinitely and is heterogeneous, cells with different sources have different genetic backgrounds, and the like. The method comprises the steps of firstly, cloning human chicken telomerase reverse transcriptase (chTERT) genes; secondly, cloning chicken telomerase (chTR) RNA genes; thirdly, constructing a reverse transcribing virus expression vector; fourthly, packaging and preparing reverse transcribing virus; and fifthly, obtaining an immortalized chicken preadipocyte system ICPA-1 and an immortalized chicken preadipocyte system ICPA-2. The method is applied to the field of building the immortalized preadipocytes of the chicken.

Methods and compositions are provided for modulating, e.g., increasing or decreasing, the expression of telomerase reverse transcriptase (TERT). In the subject methods, the binding interaction of the GC-Box 5 repressor site with a repressor protein (or protein complex including the same) is modulated to achieve the desired change in TERT expression. The subject methods and compositions find use in a variety of different applications, including the immortalization of cells, the production of reagents for use in life science research, therapeutic applications; therapeutic agent screening applications; and the like.

A compound of the formula (I), wherein: nuc is the residue of a nucleoside analogue bonded through its single hydroxy group on the cyclic or acyclic saccharide moiety, R1 is hydroxy, amino or carboxy; optionally having esterified / amide bonded thereon; a C4-C22 saturated or unsaturated, optionally substituted fatty acid or alcohol, or an aliphatic L-amino acid; R2 is the residue of an aliphatic L-amino acid; L1 is a trifunctional linker group; L2 is absent or a difunctional linker group; and pharmaceutically acceptable salts thereof have favourable pharmacological properties and are antivirally active.

The invention discloses a recombinant herpes simplex virus HSV-hTERTp_ICP4_LungCA-GFP and a corresponding diagnostic reagent kit. DNA (deoxyribonucleic acid) fragments with the lengths of 25 bp are inserted in spaces between telomerase reverse transcriptase promoters on genomes of the virus and ICP4 (infected cell protein 4) genes, GFP (green fluorescent protein) expression cassettes are inserted in ICP34.5 sites, and sequences of the DNA fragments are AGCAGGACCTCGCCCTTGGGACGAC. The recombinant herpes simplex virus and the diagnostic reagent kit have the advantages that the virus is selectively high in titer reproductive capacity, and the lung cancer cell reproduction titer of the virus is stabilized and is higher than 10<6>; synthesized random short nucleotide sequences and the genomes of the virus HSV1-hTERTp_ICP4 are subjected to site-specific recombination (between hTERTp and ICP4); the short sequences are changed, accordingly, the reproduction titer of the virus can be improved, different viruses can be selectively high in titer reproduction for different types of tumor cells, for example, certain viruses in lung cancer cells are good in reproduction, certain viruses in liver cancer cells are high in titer, and the like.

Telomerase peptides that bind MHC are disclosed. The instant application also discloses vaccines containing said peptides and methods of using said peptides to enhance a CTL response against mammalian cancer cells.

Methods and compositions are provided for modulating, e.g., increasing or decreasing, the expression of telomerase reverse transcriptase (TERT). In the subject methods, the binding interaction of the TERT Site C repressor site with a Site C repressor protein complex made up of one or more proteins is modulated to achieve the desired change in TERT expression. A feature of the subject invention is that the target Site C repressor protein complex includes a MRG15 protein. The subject methods and compositions find use in a variety of different applications, including the immortalization of cells, the production of reagents for use in life science research, therapeutic applications; therapeutic agent screening applications; and the like.

An objective of the present invention is to provide an adenovirus vector expressing a REIC / Dkk-3 protein at a high level and containing a DNA construct for expression of REIC / Dkk-3 DNA, wherein the DNA construct is prepared by ligating, from the 5′ terminal side,(i) a CMV promoter,(ii) REIC / Dkk-3 DNA,(iii) a polyA addition sequence, and(iv) enhancers prepared by linking an hTERT (Telomerase Reverse Transcriptase) enhancer, an SV40 enhancer, and a CMV enhancer in this order.

The invention discloses a cell factor fusing protein of human terminal-enzyme reverse transcriptase (hTERT) / human leucocyte 18(hIL18), which is characterized by the following: targeting tumour cell to kill; improving immune effect for dendritic cell; expanding tumour-proof scale; reducing medical cost obviously; providing the base of treating vaccine for tumour.

The present invention relates to specific immortalized avian cell lines expressing telomerase reverse transcriptase (TERT), and exhibiting distinct biologics production patterns. More particularly, the present invention relates to immortalized avian cell line capable of either amplifying Flaviviridae but not capable of amplifying Vaccinia virus strain Copenhagen (W—COP) nor Modified Vaccinia virus Ankara (MVA), or capable of amplifying both Flaviviridae and Poxyiridae. The invention further relates to the use of said immortalized avian cell lines and related methods for producing biologics, including viruses and proteins.