Abstract: The invention features lentiviral transfer vectors that include heterologous nucleic acids to be introduced into a cell. The lentiviral transfer vector may be characterized by the following features: (a) including a cytomegalovirus (CMV) promoter; (b) including a polynucleotide encoding a partial gag protein that includes a mutated INS1 inhibitory sequence that reduces restriction of nuclear export of RNA; (c) not including a polynucleotide encoding the INS2, INS3, and INS4 inhibitory sequences of gag; (d) not including an SV40 origin of replication and/or an f1 origin of replication; (e) including a cPPT sequence that contains splice site; (f) including an EF1alpha promoter with intact splice donor and acceptor sites; and (g) including hepatitis B PRE with mutation in start codon of X protein ORF.

Abstract: The present invention relates to methods and compositions for reducing the immunogenicity of chimeric Notch receptors, and specifically to transcription factors useful for controlling gene expression delivered to tissues by such chimeric Notch receptors.

Abstract: The invention features lentiviral transfer vectors that include heterologous nucleic acids to be introduced into a cell. The lentiviral transfer vector may be characterized by the following features: (a) including a cytomegalovirus (CMV) promoter; (b) including a polynucleotide encoding a partial gag protein that includes a mutated INS1 inhibitory sequence that reduces restriction of nuclear export of RNA; (c) not including a polynucleotide encoding the INS2, INS3, and INS4 inhibitory sequences of gag; (d) not including an SV40 origin of replication and/or an f1 origin of replication; (e) including a cPPT sequence that contains splice site; (f) including an EF1alpha promoter with intact splice donor and acceptor sites; and (g) including hepatitis B PRE with mutation in start codon of X protein ORF.

Abstract: The present invention relates to methods and compositions for reducing the immunogenicity of chimeric Notch receptors, and specifically to transcription factors useful for controlling gene expression delivered to tissues by such chimeric Notch receptors.

Abstract: The invention features lentiviral transfer vectors that include heterologous nucleic acids to be introduced into a cell. The lentiviral transfer vector may be characterized by the following features: (a) including a cytomegalovirus (CMV) promoter; (b) including a polynucleotide encoding a partial gag protein that includes a mutated INS1 inhibitory sequence that reduces restriction of nuclear export of RNA; (c) not including a polynucleotide encoding the INS2, INS3, and INS4 inhibitory sequences of gag; (d) not including an SV40 origin of replication and/or an f1 origin of replication; (e) including a cPPT sequence that contains splice site; (f) including an EF1alpha promoter with intact splice donor and acceptor sites; and (g) including hepatitis B PRE with mutation in start codon of X protein ORF.

Abstract: The invention provides lentiviral preparations containing a sulfonic acid buffer, such as 1,4-piperazinediethanesulfonic acid (PIPES), 2-(N-morpholino)ethanesulfonic acid (MES), and 3-morpholinopropane-1-sulfonic acid (MOPS), a sodium citrate buffer, or a phosphate buffer. The invention additionally encompasses methods of lentiviral purification as well as methods of transducing human cells.

Abstract: The invention features lentiviral transfer vectors that include heterologous nucleic acids to be introduced into a cell. The lentiviral transfer vector may be characterized by the following features: (a) including a cytomegalovirus (CMV) promoter; (b) including a polynucleotide encoding a partial gag protein that includes a mutated INS1 inhibitory sequence that reduces restriction of nuclear export of RNA; (c) not including a polynucleotide encoding the INS2, INS3, and INS4 inhibitory sequences of gag; (d) not including an SV40 origin of replication and/or an f1 origin of replication; (e) including a cPPT sequence that contains splice site; (f) including an EF1 alpha promoter with intact splice donor and acceptor sites; and (g) including hepatitis B PRE with mutation in start codon of X protein ORF.

Abstract: The invention features lentiviral transfer vectors that include heterologous nucleic acids to be introduced into a cell. The lentiviral transfer vector may be characterized by the following features: (a) including a cytomegalovirus (CMV) promoter; (b) including a polynucleotide encoding a partial gag protein that includes a mutated INS1 inhibitory sequence that reduces restriction of nuclear export of RNA; (c) not including a polynucleotide encoding the INS2, INS3, and INS4 inhibitory sequences of gag; (d) not including an SV40 origin of replication and/or an f1 origin of replication; (e) including a cPPT sequence that contains splice site; (f) including an EF1 alpha promoter with intact splice donor and acceptor sites; and (g) including hepatitis B PRE with mutation in start codon of X protein ORF.

Abstract: The present invention relates to methods and compositions for reducing the immunogenicity of chimeric Notch receptors, and specifically to transcription factors useful for controlling gene expression delivered to tissues by such chimeric Notch receptors.

Abstract: The invention provides lentiviral preparations containing a sulfonic acid buffer, such as 1,4-piperazinediethanesulfonic acid (PIPES), 2-(N-morpholino)ethanesulfonic acid (MES), and 3-morpholinopropane-1-sulfonic acid (MOPS), a sodium citrate buffer, or a phosphate buffer. The invention additionally encompasses methods of lentiviral purification as well as methods of transducing human cells.

Abstract: The present invention provides for lentiviral vectors for vaccine delivery comprising a 5? long terminal repeat (LTR) and a 3? LTR, an integrated nucleic acid sequence, wherein the integrated nucleic acid sequence is expressed by the 5? LTR; and a nucleic acid sequence encoding functional REV coding sequence and a rev response element (RRE)-containing sequence, wherein the RRE-containing sequence is located upstream of the REV coding sequence, and wherein transcription of said first nucleic acid sequence and said second nucleic acid sequence is driven by said 5? LTR. Also provided for are pharmaceutical compositions, methods of making and using the lentiviral vectors of the present invention.

Abstract: The invention provides novel processes for manufacturing autologous T cells, transducing T cells and expanding the transduced T cell population.

Abstract: The invention relates to a method of increasing vector transduction in target cells. The invention provides for the recombinant engineering of a packaging cell line to be capable of expressing one or more membrane proteins which facilitate binding to, and activation of, a target cell. The invention also provides for recombinant engineering of a cell that endogenously expresses one or more such membrane proteins into a packaging cell line. A vector packaged into viral particles via use of such cell lines would comprise an outer envelope containing these proteins. The particles would be specifically suited for binding and targeting to a target cell to facilitate transduction thereof with the vector. The target cell may also be simultaneously activated (stimulated) by the packaged vector in the absence of exogenously supplied stimulatory molecules.

Abstract: The present invention provides a recombinant transcription unit capable of producing an RNA transcript of a predetermined size comprising a regulatory sequence operably linked to a nucleotide sequence comprising a transcribed region such that the transcription of said transcribed region is controlled by said regulatory sequence. The transcribed region comprises a region that encodes for a viral sequence, and a non-coding region downstream of the region encoding for said viral sequence, wherein the non-coding region comprises a nucleotide sequence encoding a cis-acting ribozyme. Methods of using the recombinant transcription unit, and cells containing vectors comprising the recombinant transcription unit are also disclosed.

Abstract: The present invention provides methods, as well as compositions related thereto, for the efficient transduction of cells using viral vectors. The efficiency of transduction is increased by contacting the cell to be transduced with one or more molecules that bind the cell surface. Contact with a cell surface binding molecule may occur before, after, or simultaneously with contact between the viral vector and the cell. The transduced vectors may be constructed to express a gene of interest, permitting the transduced cells to be used as therapeutic and prophylactic agents.

Abstract: The present invention provides methods, as well as compositions related thereto, for the efficient transduction of cells using viral vectors. The efficiency of transduction is increased by contacting the cell to be transduced with one or more molecules that bind the cell surface. Contact with a cell surface binding molecule may occur before, after, or simultaneously with contact between the viral vector and the cell. The transduced vectors may be constructed to express a gene of interest, permitting the transduced cells to be used as therapeutic and prophylactic agents.

Abstract: The present invention discloses compositions and methods of using intracellular delivery vehicles for delivery and transfection of DNA, RNA, polypeptides, genes, proteins, drugs and biologically active agents into cells in vitro and in vivo. The vehicle comprises a mixture of a liposome and a polypeptide lacking specificity for cellular receptors. In another embodiment, a method for intracellular delivery of biologically active agents comprising combining a non-receptor-binding protein and a liposome, incubating the mixture for a period of time, adding the biologically active agent, incubating again, and finally, introducing the resulting mixture to the cell. Preferably, the liposome is a cationic liposome. The charge ratio of cationic liposome to DNA can effectively be varied from 2:1 to 1:2. Preferably, the non-receptor-binding protein is the serum albumin of the animal source of the cell to be transfected.