54 results about "Pol genes" patented technology

Non-infectious, retrovirus-like particles comprise an assembly of an env gene product, a pol gene product and a gag gene product contain an antigenic marker which is non-retroviral or non-HIV retroviral. In one embodiment, the marker comprises an amino acid sequence containing an epitope inserted into the gag gene product at an antigenically-active insertion site. In another embodiment, the marker comprises an antigenic anchor sequence operatively connected to the env gene product replacing endogenous anchoring function. The corresponding nucleic acid molecules are described. The non-infectious, retrovirus-like particles have utility in in vivo administration including to humans and in diagnosis. The presence of the antigenic marker enables recognition that antiserum containing anti-retroviral antibodies has been generated by exposure to the non-infectious retrovirus-like particles by testing for antibodies specific to the antigenic marker.

Novel packaging cell lines which produce recombinant retrovirus, free of detectable helper-virus are disclosed. Also disclosed are methods of making the cell lines and methods of producing recombinant retroviruses from the cell lines. Retroviruses produced by the cell lines include lentiviruses, such as HIV, capable of transfering heterologous DNA to a wide range of non-dividing cells. The packaging cells contain at least three vectors which collectively encode retroviral gag, pol, and env proteins, wherein the gag and pol genes are separated, in part, onto two or more different vectors. This is made possible by fusing Vpr or Vpx to pol proteins separated from gag so that the proteins are targeted to assembling virions. Among other advantages, the packaging cells provide the benefit of increased safety when used in human gene therapy by virtually eliminating the possibility of molecular recombination leading to production of replication-competant helper virus.

Non-infectious, retrovirus-like particles comprise an assembly of an env gene product, a pol gene product and a gag gene product contain an antigenic marker which is non-retroviral or non-HIV retroviral. In one embodiment, the marker comprises an amino acid sequence containing an epitope inserted into the gag gene product at an antigenically-active insertion site. In another embodiment, the marker comprises an antigenic anchor sequence operatively connected to the env gene product replacing endogenous anchoring function. The corresponding nucleic acid molecules are described. The non-infectious, retrovirus-like particles have utility in in vivo administration including to humans and in diagnosis. The presence of the antigenic marker enables recognition that antiserum containing anti-retroviral antibodies has been generated by exposure to the non-infectious retrovirus-like particles by testing for antibodies specific to the antigenic marker.

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 in vivo administration including to humans and in diagnosis.

The present invention provides a process for preparing a retrovirus to be expressed at a high titer by specifically transferring a desired foreign gene into target cells. A pseudotyped retrovirus vector having a high titer can be prepared by transferring a DNA construction wherein a promoter, an loxP sequence, a VSV-G gene and a polyA addition signal are arranged in this order is transferred into cells carrying the retrovirus gag and pol gene expression systems, and then transferring a retrovirus vector containing the desired foreign gene thereinto, followed by treatment with a recombinase.

The invention discloses a triple minRNA for resisting virus infection of aids and the construction method thereof. The sequences of the triple minRNA are respectively designed according to gene sequences of ccr5, pol and vif and combined to be miRNA oligomeric and single strand DNA, then double-strand DNA is formed through annealing, then double-strand DNA is respectively inserted into the expression vector of pcDNA<TM> 6.2-GW / EmGFPmiR of miRNA to built the mi RNA expression plasmid, and the jamming effect of mi RNA expression plasmid to target gene can be detected through a Qpcr method. A miRNA string with the best gene interference effect to ccr5, vif and pol is inserted into a carrier of pcDNA 6.2 <TM>-GW / EmGFP, so as to obtain a series-wound interference vector of pcDNA6.2<TM>-GW / EmGFP-ccr5-pol-vif, and then the miR-ccr5-pol-vif in the series-wound interference vector is transferred to a lentiviral vector pLenti6.3 / V5-DEST so as to construct pLenti6.3 / V5-DEST miR-ccr5-pol-vif. The recombinant lentiviral vector can product three miRNA at the same time to cause interference effects to gene ccr5, gene pol and gene vif, so as to prevent the gene ccr5, the gene pol and the gene vif from expressing corresponding protein. Therefore, the effect of controlling Hiv infection and replication can be achieved.

The present invention is directed toward methods for the production of non-infectious, replication-deficient, immunogenic human immunodeficiency virus (HIV)-like particles. These particles are prepared from a recombinant expression vector comprising a heterologous promoter operatively connected to a DNA molecule comprising a modified HIV genome devoid of the long terminal repeat (LTR) regulatory regions but containing at least the gag and pol genes in their natural genomic arrangement. This vector is introduced into mammalian cells to produce the particles of interest. These particles should prove useful in a number of diagnostic, virologic, and immunologic applications.

The present invention provides a process for preparing a retrovirus to be expressed at a high titer by specifically transferring a desired foreign gene into target cells. A pseudotyped retrovirus vector having a high titer can be prepared by transferring a DNA construction wherein a promoter, an loxP sequence, a VSV-G gene and a polyA addition signal are arranged in this order is transferred into cells carrying the retrovirus gag and pol gene expression systems, and then transferring a retrovirus vector containing the desired foreign gene thereinto, followed by the treatment with a recombinase.

The present invention relates to a method for obtaining stable pseudotyped lentiviral particles including a heterologous gene of interest, comprising the following steps: a) transfecting at least one plasmid in appropriate cell lines, wherein said at least one plasmid comprises the gene of interest, the rev. gag and pol genes, and a sequence coding for an ERV syncytin, wherein the rev, gag and pol genes are retroviral genes; b) incubating the transfected cells obtained in a), so that they produce the stable pseudotyped lentiviral particles in the supernatant; and c) harvesting and concentrating the stable lentiviral particles obtained in b). The present invention also relates to a method to transduce immune cells using lentiviral vectors pseudotyped with an ERV syncytin glycoprotein. The method can be performed on non-stimulated blood cells or on cells stimulated briefly with IL7, and the cells can be expanded. The stable pseudotyped lentiviral particles obtained are particularly useful in gene therapy.

The present invention relates to non-replicative recombinant retrovirus packaging cells able to grow in suspension in a serum-free medium. In particular, the present invention relates to a human embryonic 293SF-based cell line stably expressing gag and pol gene products from the murine Moloney leukemia virus (MLV) and either the feline RD114 env gene, the gibbon ape leukemia virus (GLV) env gene, or the amphotropic 4070Aenv gene. This particular combination allows the production of high titer of non-replicative retrovirus pseudotyped and prevents the recombination of plasmids. The recombinant retroviruses produced from these cells are safer and easier to produce for clinical use in gene therapy.

The present invention relates to a method for mutation analysis of the HIV pol gene of HIV virions comprising amplifying virion RNA or DNA via nested PCR using outer primers as represented in SEQ ID No. 1 and 2, amplifying said PCR product via nested PCR using a 5′ and 3′ primer chosen from the inner primers SEQ ID No. 3, 4, 5, and 6, and sequencing this secondary obtained PCR product using at least one sequencing primer chosen from any of SEQ ID No. 7 to 12 or variants thereof. In the alternative, at least one secondary sequencing primer may be used chosen from any of SEQ ID No. 13 to 24. The benefit of the sequences present in the invention resides in the fact that, with the aid of the oligonucleotides, the sequences of all presently known HIV subtypes and all mutations of the pol gene presently known to yield resistance towards antiretroviral therapy can be determined. The present invention also relates to kits for performing such a method as well as primers for performing the same.

The invention discloses a strawberry reverse transcription transposon gene sequence and a transcription characteristic analysis result. A full length of a gene DNA sequence is 5386 bp, and is named as FaREII, the gene sequence has a Ty1-copia reverse transcription transposon gene characteristic sequence, and is arranged according to gag and pol genes, wherein pol gene contains the characteristic motifs of GAG, PR, IN, RT and RH genes, a long terminal repeat sequence of 393 bp is positioned at two ends of the gene, and the 393 bp is a typical Ty1-copia reverse transcription transposon. According to the invention, the transcription of FaREII can be happened under hormone stress through Nothern Blot.