55 results about "Long terminal repeat" patented technology

Non-infectious, retrovirus-like particles contain mutations to reduce gag-dependent RNA-packaging of the gag gene product, eliminate reverse transcriptase activity of the pol gene product, eliminate integrase activity of the pol gene product and eliminate RNase H activity of the pol gene product through genetic manipulation of the gag and pol genes. The corresponding nucleic acid molecules are described. The non-infectious, retrovirus-like particles have utility in diagnosis.

A method of inactivating a proviral DNA integrated into the genome of a host cell latently infected with a retrovirus by treating the host cell with a composition comprising a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease, and two or more different guide RNAs (gRNAs), wherein each of the at least two gRNAs is complementary to a different target nucleic acid sequence in a long terminal repeat (LTR) in the proviral DNA, and inactivating the proviral DNA. A composition for use in inactivating a proviral DNA integrated into the genome of a host cell latently infected with a retrovirus including isolated nucleic acid sequences comprising a CRISPR-associated endonuclease and a guide RNA, wherein the guide RNA is complementary to a target sequence in a human immunodeficiency virus.

This invention provides transcriptionally-activated AAV ITRs (inverted terminal repeats) which are small and transcriptionally active and uses thereof to optimize the expression of relatively large transgenes packaged in recombinant AAV vectors.

Non-infectious, non-replicating immunogenic HIV-like particles are produced by stable longn-term constitutive expression in mammalian cells by eliminating elements toxic to the mammalian cells. An expression vector contains a nucleic acid molecule comprising a modified HIV genome devoid of long terminal repeats and wherein Tat and vpr sequences are functionally disabled and a constitutive promoter operatively connected to the modified HIV genome for constitutive expression of the modified genome to produce the HIV-like particles.

The present invention provides an isolated nucleic acid comprising a single retroviral LTR, a polypurine tract, a packaging signal, a primer binding site and a rev responsive element. Further provided is an isolated nucleic acid comprising a heterologous nucleotide sequence, a single retroviral long terminal repeat (LTR), a packaging signal, a rev responsive element, a polypurine tract, a eukaryotic promoter, a primer binding site, a bacterial origin of replication and a bacterial selection marker. In addition, the present invention provides an isolated nucleic acid comprising a 5′ retroviral LTR and a 3′ retroviral LTR, a heterologous nucleotide sequence, a packaging signal, a rev responsive element, a polypurine tract, a eukaryotic promoter, a primer binding site, a bacterial origin of replication and a bacterial selection marker cassette, wherein the bacterial origin of replication and bacterial selection marker are located between the two LTRs.

The present invention relates to a method for increasing retrovirus transcription in an infected eukaryotic cell, comprising increasing Wnt pathway signaling in said cell such that transcription of said retrovirus is increased. Also, the present invention relates to a method of treating a subject infected with a retrovirus, said method comprising administering to said subject an activator of the Wnt pathway in an amount that increased Wnt pathway signaling in resting memory CD4+ T cells of said subject such that the retroviral long terminal repeat (LTR) in said cells is activated or de-repressed.

The invention relates to a gene detection kit against human immunodeficiency virus-1 (HIV-1). The gene detection kit comprises a pair of amplification primers, a capture probe, a detection probe, an NASBA (National Association of State Boards of Accountancy) amplification reagent and an NASBA product detection reagent. The gene detection kit provided by the invention designs the primers against the long terminal repeat sequence of the HIV-1, establishes an NASBA fast detection method and is suitable for early diagnosis of HIV-1 infection.

The present invention relates to the expression and screening of genomic DNA sequences encoding uncharacterized genes and proteins. The present invention provides systems utilizing unique features of retroviral replication to analyze uncharacterized genes derived from genomic DNA samples. In preferred embodiments, a segment of genomic DNA is inserted between 5′ and 3′ viral long terminal repeats (LTRs) in a vector (e.g., a plasmid, cosmid, or artificial chromosome vector). The resulting vector (or library of vectors containing a plurality of independent genomic sequences) is then introduced into a retroviral packaging cell. The resulting provirus or proteins expression from the provirus are then analyzed.

The present invention discloses sequences having a structure and sequence homology to lox-P recombinase target site corresponding to a highly conserved region within the Long Terminal Repeats (LTR) of HIV and to the use thereof for the identification of recombinase enzymes useful in treating HIV-1.

The invention particularly relates to a high-infection quasi-influenza virus system which is mainly characterized in that carriers of LTR(Long Terminal Repeat) containing HIV(Human Immunodeficiency Virus)-1, promoters, gag-pol and other elements and high-infection influenza virus membrane protein expression plasmids are subjected to co-transfection and packaging to form quasi-influenza viruses which have infection characteristics and antigenicity of high-infection influenza virus membrane protein. The system is used as a convenient and safe platform for researches on the physiological characteristics of highly pathogenic influenza virus membrane protein and researches on neutralizing antibodies. The system is used as an efficient gene introduction carrier, a target gene can be inserted into a gene-containing system reconstructed through packaging signals of the HIV-1, and the target gene can be introduced into various cells through quasi-virus infection according to retrovirus integration rules. After the cells are infected by quasi-viruses, reporter genes are integrated and expressed through gag-pol protein of the HIV-1, and the inhibition to the gag-pol function by medicaments can be reflected through the expression of the reporter genes, thus the system can be used as an evaluation platform for gag-pol medicaments.

The present disclosure provides for recombinant nucleic acids, and cells and virions comprising the recombinant nucleic acids, that can be used to identify, isolate, and/or purify cells latently infected with immunodeficiency virus. A subject recombinant nucleic acid includes (a) a first nucleotide sequence encoding a first reporter polypeptide that produces a first detectable signal, where the first nucleotide sequence is operably linked to an immunodeficiency virus promoter and is translated as an early gene; and (b) a second nucleotide sequence encoding a second reporter polypeptide that produces a second detectable signal that is distinguishable from the first detectable signal, where the second nucleotide sequence is operably linked to a non-immunodeficiency virus promoter. In some aspects, the first and second nucleotide sequences are both positioned between a shared 5′ long terminal repeat (LTR) and a shared 3′ LTR. Also provided are related methods.

A method of inactivating a proviral DNA integrated into the genome of a host cell latently infected with a retrovirus by treating the host cell with a composition comprising a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease, and two or more different guide RNAs (gRNAs), wherein each of the at least two gRNAs is complementary to a different target nucleic acid sequence in a long terminal repeat (LTR) in the proviral DNA, and inactivating the proviral DNA. A composition for use in inactivating a proviral DNA integrated into the genome of a host cell latently infected with a retrovirus including isolated nucleic acid sequences comprising a CRISPR-associated endonuclease and a guide RNA, wherein the guide RNA is complementary to a target sequence in a human immunodeficiency virus.

An immunogenic retrovirus-like particle which is non-infectious and non-replicating and which is useful as a candidate vaccine component against retroviral infections, including AIDS and ATLL, is produced by genetic engineering. A DNA molecule comprising a retroviral genome devoid of long terminal repeats is incorporated into an expression vector, which is introduced into mammalian cells for expression of the retrovirus-like particle.

A method for monitoring ARV resistance, to determine viral fitness, and to forecast possible drug failure utilizes two nucleic acid sequences. One nucleic acid includes a retroviral nucleic acid devoid of at least a majority of the sequence for one of the two long terminal repeat regions. A second nucleic acid, includes a retroviral nucleic acid sequence devoid of the sequences encoding an envelope gene and the second long terminal repeat region of the retrovirus. The method allows the rapid cloning of an amplicon into an HIV-1 genome vector through recombination/gap repair in organisms such as yeast. The vectors can be directly passed to a mammalian cell line which has been specifically engineered to produce replication competent HIV-1 particles. The susceptibility of an isolate to any of several ARVs, i.e. PRIs, NRTIs, NNRTIs, T20, as well as entry and integrase inhibitors in developmentlclinical trials, may be tested.

There is disclosed a retroviral vector comprising a primer binding site, a long terminal repeat and an RNA packaging sequence, wherein the RNA packaging sequence is located 3′ of the long terminal repeat and no long terminal repeat is located 3′ of the RNA packaging sequence such that reverse transcription initiated at the primer binding site does not lead to reverse transcription of the RNA packaging sequence into vector DNA in a target cell. Also described is a host cell, a virion, a pharmaceutical composition, a method and uses including or involving the vector described above. Further, a cell or transgenic animal produced by using the vector is also described.

The present invention relates to a method for preparing an expression vector encoding a tailored recombinase, which tailored recombinase is capable of recombining asymmetric target sequences within the long terminal repeat (LTR) of proviral DNA of a plurality of retrovirus strains inserted into the genome of a host cell, as well as to the obtained expression vector, cells transfected with this, expressed recombinase and pharmaceutical compositions comprising the expression vector, cells and/or recombinase. Pharmaceutical compositions are useful, e.g., in treatment and/or prevention of retrovirus infection. In particular, asymmetric target sequences present in a plurality of HIV strains are disclosed, as well as tailored recombinases capable of combining these sequences (Tre 3.0 and 4.0) and expression vectors encoding them.

The application discloses a recombinant lentivirus vector for treating beta-globulin afunction, a preparation method and application. A gene element of the recombinant lentivirus vector disclosed by the application is composed of a left side lentivirus long terminal repeat, a lentivirus counter-response element, a center poly-purine sequence, a gene locus regulating and controlling sequence of a beta-globulin gene, a beta-globulin gene, of which 87-th threonine is mutated into glutamine, and a right side lentivirus long terminal repeat which are connected in order. According to the recombinant lentivirus vector disclosed by the application, through optimizing and improving the gene element, the gene transduction efficiency is increased, and the beta-globulin gene can be efficiently expressed during erythroid differentiation of hemopoietic stem cells, so that the consumption of viruses can be effectively lowered, and the treatment cost is reduced while the safety is improved. The recombinant lentivirus vector disclosed by the application provides a novel treatment tool and scheme for beta-thalassemia and sickle cell anemia caused by the beta-globulin afunction.