1333 results about "Reverse transcriptase" patented technology

The present invention provides a method for determination of the identity of at least one nucleotide in a RNA-molecule comprising the steps of: (i) providing the RNA-molecule, an oligonucleotide primer binding to a predetermined position of the RNA molecule, a reverse transcriptase, deoxynucleotides and other necessary reagents, in a reaction vessel; (ii) performing a primer extension reaction, whereby the oligonucleotide primer is extended on the RNA-molecule through incorporation of at least one deoxynucleotide by the action of a reverse transcriptase, resulting in the release of a PPi molecule only upon incorporation of a deoxynucleotide; and (iii) detecting the presence or absence of incorporation, thereby indicating the nucleotide identity of the RNA molecule in the relevant position. In a preferred embodiment, the sequencing of the invention is coupled to the Pyrosequencing™ reaction. A variant of the method employs incorporation of modified nucleotides, with an optionally cleavable linker arm to which is attached a label.

The present invention provides methods and compositions for protein stabilization, particularly the stabilization of polymerases in aqueous solutions with cationic surfactants. The present invention further provides cationic surfactants, including polyethoxylated amines, that stabilize thermostable and thermolabile enzymes in solution. These surfactants stabilize the activity of various enzymes, including thermostable DNA polymerases, thermolabile DNA polymerases and reverse transcriptases.

2′ and 3′-Prodrugs of 1′, 2′, 3′ or 4′-branched β-D or β-L nucleosides, or their pharmaceutically acceptable salts and derivatives are described, which are useful in the prevention and treatment of Flaviviridae infections and other related conditions. These modified nucleosides provide superior results against flaviviruses and pestiviruses, including hepatitis C virus and viruses generally that replicate through an RNA dependent RNA reverse transcriptase. Compounds, compositions, methods and uses are provided for the treatment of Flaviviridae infection, including HCV infection, that include the administration of an effective amount of the prodrugs of the present invention, or their pharmaceutically acceptable salts or derivatives. These drugs may optionally be administered in combination or alteration with further anti-viral agents to prevent or treat Flaviviridae infections and other related conditions.

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.

The present invention describes methods for the characterization of mRNA molecules during mRNA production. Characterizing mRNA includes processes such as oligonucleotide mapping, reverse transcriptase sequencing, charge distribution analysis, and detection of RNA impurities. Oligonucleotide mapping includes using an RNase to digest antisense duplexes from an RNA transcript, and then subjecting the digested RNA to reverse phase HPLC, anion exchange HPLC, and/or mass spectrometry analysis. Reverse transcriptase sequencing involves reverse transcription of an RNA transcript followed by DNA sequencing. Charge distribution analysis can comprise procedures such as anion exchange HPLC, or capillary electrophoresis. Detection of impurities includes detecting short mRNA transcripts, RNA-RNA hybrids, and RNA-DNA hybrids.

Compounds having the structure:

are HIV reverse transcriptase inhibitors, wherein A, X, Y, Z, R¹ and R² are defined herein. The compounds and their pharmaceutically acceptable salts are useful in the inhibition of HIV reverse transcriptase, the prophylaxis and treatment of infection by HIV and in the prophylaxis, delay in the onset, and treatment of AIDS. The compounds and their salts can be employed as ingredients in pharmaceutical compositions, optionally in combination with other antivirals, immunomodulators, antibiotics or vaccines.

An anti-HIV agent containing, as an active ingredient, a 4-oxoquinoline compound represented by the following formula [I]

wherein each symbol is as defined in the specification, or a pharmaceutically acceptable salt thereof. The compound of the present invention has HIV integrase inhibitory action and is useful as an anti-HIV agent for the prophylaxis or therapy of AIDS. Moreover, by a combined use with other anti-HIV agents such as protease inhibitors, reverse transcriptase inhibitors and the like, the compound can become a more effective anti-HIV agent. Since the compound has high inhibitory activity specific for integrases, it can provide a safe pharmaceutical agent with a fewer side effects for human.

The present invention provides a therapeutic compound of formula (I) and their pharmaceutically acceptable salts for the prevention and treatment of lipodystrophy caused because of HIV infection or combination therapy of HIV-1 protease inhibitors (Pis) and/or reverse transcriptase inhibitors (nRTIs) by neutralizing lipohypertrophy, lipoatrophy and metabolic abnormalities in HIV patient.

A series of S-triazolyl α-mercaptoacetanilides having general structure (1) are provided, where Q is CO₂H, CONR₂, SO₃H, or SO₂NR₂. The compounds inhibit several variants of the reverse transcriptase of HIV, and are useful in the treatment of HIV infections.

Compounds and compositions for the transient expression of exogenous telomerase activity in a cell are provided. The compounds and compositions, which relate to a ribonucleic acid coding for a telomerase reverse transcriptase, are useful in the extension of telomeres in cells needing such treatment. Such cells include, for example, cells that contain shortened telomeres and cells from subjects that may benefit from telomere extension, for example subjects that suffer from, or are at risk of suffering from, age-related or other illnesses. Also provided are methods of extending telomeres through the administration of the provided compounds and compositions to animal cells, either in vitro or in vivo, and kits including the compounds or compositions and instructions for use.

The present invention pertains to a method to isolate, separate, enrich or amplify a targeted nucleotide polymer such as mRNA through selective reverse transcription of the targeted polymer into cDNA from a sample comprising of chemically identical or similar polynucleotide polymers such as rRNA. The enrichment of the targeted nucleic acid such as mRNA is accomplished by blocking the reverse transcription of undesired rRNA while allowing unrestricted reverse transcription of the targeted polymer. The invention also embodies that the cleavage of the non-targeted nucleic acid such as rRNA bound to an oligonucleotide through enzymatic activity (RNase H). The invention further embodies methods and kits to accomplish the utility of the invention through the following steps 1) 3′ tailing of chemically identical or similar nucleotide polymers in a sample that includes bacterial mRNA 2) a 3′ tail capable of binding to a oligo-dN primer 3) at least one oligonucleotide capable of preventing the extension of oligo-dN bound to at least one non-targeted nucleotide polymers by a DNA polymerase such as a reverse transcriptase without restricting conversion of bacterial mRNA into cDNA 4) where the non-targeted molecule is prevented as a template for cDNA synthesis by enzymatic cleavage (RNase H) of template (rRNA)-oligonucleotide hybrid 5) where the reverse transcriptase is physically blocked by the oligonucleotide bound to the non-targeted nucleic acids such as rRNA 5) purification of the selectively transcribed cDNA. In further embodiments of the present invention, methods and composition to enable the study of bacterial transcriptomics-an analysis of genes expressed by a bacterial infection of a host, an isolated bacterial culture or a bacterial community, such as recovered from soil, intestine, mouth, biofilm, water etc are also included for use in DNA-chip or sequencing analyses.

A method for synthesizing cDNA possessing a consecutive sequence starting with a nucleotide adjacent to a cap structure of mRNA, which comprises (i) a process for annealing a double-stranded DNA primer and an RNA mixture containing mRNA possessing a cap structure, (ii) a process for preparing a conjugate of an mRNA/cDNA heteroduplex and a double-stranded DNA primer by synthesizing the first-strand cDNA primed with the double-stranded DNA primer using reverse transcriptase, and (iii) a process for circularizing the conjugate of the mRNA/cDNA heteroduplex and the double-stranded DNA primer by joining the 3' and 5' ends of the DNA strand containing cDNA using ligase. This method enables us to efficiently synthesize a full-length cDNA possessing a consecutive sequence starting with a transcription-start-site nucleotide from a small amount of RNA by small processes.

The present invention is directed to a novel variant of human telomerase reverse transcriptase (S16AhTERT), which displays properties distinct from those of wildtype telomerase reverse transcriptase. Accordingly, the amino acid sequence of S16AhTERT and nucleic acid sequences encoding same are presented herein, as are methods of use thereof.

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.

Current methods for detecting influenza A subtype H5 virus, for example cell culture, haemagglutination-inhibition, fluorescent antibody and enzyme immunoassay, and reverse transcriptase polymerase chain reaction (RT-PCR) may have the disadvantages of low sensitivity and low specificity. Furthermore, such methods are relatively difficult to use, and may not be suitable for routine detection on a daily basis. The kit for detecting H5 virus of this invention may provide a user-friendly alternative that is relatively more sensitive and specific to H5 virus. The detection kit utilizes two specially designed primers A and B for the replication of H5 virus, and a specific capture probe for immobilizing the amplified viral RNA. An additional primer C is also designed for the detection of pathogenic H5 virus. The detection of H5 virus by the detection kit may be accomplished within one day if desired.

There is provided inter alia use of 2′,3′-dideoxy-3′-hydroxymethylcytosine or a prodrug or salt thereof in the manufacture of a medicament for the treatment of HIV infection wherein the reverse transcriptase of the HIV bears at least one mutation that allows an obligate chain terminating nucleoside- or nucleotide phosphate to be excised from the nascent DNA strand by ATP- or pyrophosphate-mediated excision.