Abstract: The present invention enables simultaneous and stable expression of a plurality of foreign genes by using a stealthy RNA gene expression system that is a complex that does not activate the innate immune mechanism and is formed from an RNA-dependent RNA polymerase, a single-strand RNA binding protein, and negative-sense single-strand RNAs including the following (1) to (8): (1) a target RNA sequence that codes for any protein or functional RNA; (2) an RNA sequence forming a noncoding region and derived from mRNA expressed in animal cells; (3) a transcription initiation signal sequence recognized by the RNA-dependent RNA polymerase; (4) a transcription termination signal sequence recognized by the polymerase; (5) an RNA sequence containing a replication origin recognized by the polymerase; (6) an RNA sequence that codes for the polymerase and of which codons are optimized for the species from which an introduction target cell is derived; (7) an RNA sequence that codes for a protein for regulating the activity o

Abstract: The present invention enables simultaneous and stable expression of a plurality of foreign genes by using a stealthy RNA gene expression system that is a complex that does not activate the innate immune mechanism and is formed from an RNA-dependent RNA polymerase, a single-strand RNA binding protein, and negative-sense single-strand RNAs including the following (1) to (8): (1) a target RNA sequence that codes for any protein or functional RNA; (2) an RNA sequence forming a noncoding region and derived from mRNA expressed in animal cells; (3) a transcription initiation signal sequence recognized by the RNA-dependent RNA polymerase; (4) a transcription termination signal sequence recognized by the polymerase; (5) an RNA sequence containing a replication origin recognized by the polymerase; (6) an RNA sequence that codes for the polymerase and of which codons are optimized for the species from which an introduction target cell is derived; (7) an RNA sequence that codes for a protein for regulating the activity o

Abstract: An isolated RNA molecule includes an artificial microRNA precursor comprising in the 5??3? direction: a first terminal oligonucleotide consisting of AGGCCR (SEQ ID NO: 1) or a nucleotide sequence in which one to three nucleotides in SEQ ID NO: 1 are substituted; a passenger strand oligonucleotide; a first central oligonucleotide consisting of CYG (SEQ ID NO: 2); a second central oligonucleotide consisting of a nucleotide sequence having at least 70% homology with UUGAAUAKAAAU (SEQ ID NO: 3); a third central oligonucleotide consisting of YGG (SEQ ID NO: 4); a guide strand oligonucleotide; and a second terminal oligonucleotide consisting of UGGAYYK (SEQ ID NO: 5) or a nucleotide sequence in which one to three nucleotides in SEQ ID NO: 5 are substituted.

Abstract: The present invention enables simultaneous and stable expression of a plurality of foreign genes by using a stealthy RNA gene expression system that is a complex that does not activate the innate immune mechanism and is formed from an RNA-dependent RNA polymerase, a single-strand RNA binding protein, and negative-sense single-strand RNAs including the following (1) to (8) : (1) a target RNA sequence that codes for any protein or functional RNA; (2) an RNA sequence forming a noncoding region and derived from mRNA expressed in animal cells; (3) a transcription initiation signal sequence recognized by the RNA-dependent RNA polymerase; (4) a transcription termination signal sequence recognized by the polymerase; (5) an RNA sequence containing a replication origin recognized by the polymerase; (6) an RNA sequence that codes for the polymerase and of which codons are optimized for the species from which an introduction target cell is derived; (7) an RNA sequence that codes for a protein for regulating the activity

Abstract: The present invention enables simultaneous and stable expression of a plurality of foreign genes by using a stealthy RNA gene expression system that is a complex that does not activate the innate immune mechanism and is formed from an RNA-dependent RNA polymerase, a single-strand RNA binding protein, and negative-sense single-strand RNAs including the following (1) to (8): (1) a target RNA sequence that codes for any protein or functional RNA; (2) an RNA sequence forming a noncoding region and derived from mRNA expressed in animal cells; (3) a transcription initiation signal sequence recognized by the RNA-dependent RNA polymerase; (4) a transcription termination signal sequence recognized by the polymerase; (5) an RNA sequence containing a replication origin recognized by the polymerase; (6) an RNA sequence that codes for the polymerase and of which codons are optimized for the species from which an introduction target cell is derived; (7) an RNA sequence that codes for a protein for regulating the activity o

Abstract: Simultaneous expression of a plurality of foreign genes by using a stealthy RNA gene expression system that is a complex that does not activate the innate immune mechanism and is formed from an RNA-dependent RNA polymerase, a single-strand RNA binding protein, and negative-sense single-strand RNAs including the following (1) to (8): (1) a target RNA sequence that codes for any protein or functional RNA; (2) an RNA sequence forming a noncoding region and derived from mRNA; (3) a transcription initiation signal sequence recognized by the RNA-dependent RNA polymerase; (4) a transcription termination signal sequence recognized by the polymerase; (5) an RNA sequence containing a replication origin recognized by the polymerase; (6) an RNA sequence that codes for the polymerase; (7) an RNA sequence that codes for a protein for regulating the activity of the polymerase; and (8) an RNA sequence that codes for the single-strand RNA binding protein.

Abstract: Simultaneous expression of a plurality of foreign genes by using a stealthy RNA gene expression system that is a complex that does not activate the innate immune mechanism and is formed from an RNA-dependent RNA polymerase, a single-strand RNA binding protein, and negative-sense single-strand RNAs including the following (1) to (8): (1) a target RNA sequence that codes for any protein or functional RNA; (2) an RNA sequence forming a noncoding region and derived from mRNA; (3) a transcription initiation signal sequence recognized by the RNA-dependent RNA polymerase; (4) a transcription termination signal sequence recognized by the polymerase; (5) an RNA sequence containing a replication origin recognized by the polymerase; (6) an RNA sequence that codes for the polymerase; (7) an RNA sequence that codes for a protein for regulating the activity of the polymerase; and (8) an RNA sequence that codes for the single-strand RNA binding protein.

Abstract: A reprogramming gene-loaded Sendai viral vector comprising Sendai virus genes and reprogramming genes, wherein the Sendai virus genes include an NP gene, P/C gene, M gene, F gene, HN gene and L gene, wherein each of the M gene, the F gene and the FIN gene is from a Sendai virus strain Cl.151-derived gene and wherein at least one of the M gene, the F gene and the HN gene is functionally deleted and the L gene encodes the amino-acid sequence of the L protein in which the amino-acid residue at position 1618 is valine and a method of producing the same.

Abstract: A persistently infective virus vector is produced by using a gene so modified as to encode an amino acid sequence including a valine substituted for an amino acid residue at position-1618 in the amino acid sequence for an L protein of a persistently non-infective Sendai virus. A non-transmissible, persistently infective virus vector is also produced by defecting or deleting at least one of M gene, F gene, and HN gene. These virus vectors have no cytotoxicity, can achieve the sustained gene expression over a long period of time, is safe, and is therefore useful.

Abstract: Stem cell reprogramming genes cloned into a single sustained expression-type Sendai viral vector are shown to reprogram differentiated somatic cells into induced pluripotent stem (iPS) cells without integration of vector sequences into the host cell's genome. The genes are transduced into normal differentiated somatic cells via infection with recombinant Sendai virus. After expression of the reprogramming genes and subsequent induction of pluripotency, the vector genome RNA including the reprogramming genes is removed from the cell to establish an iPS cell that is genetically identical to the parent somatic differentiated cell thus reducing the risk of tumorigenic transformation caused by random integration of vector sequences into the host genome. The method promises to provide safe, autologous iPS cells that can be used for human cell replacement and regeneration therapeutic applications.

Abstract: A reprogramming gene-loaded Sendai viral vector comprising Sendai virus genes and reprogramming genes, wherein the Sendai virus genes include an NP gene, P/C gene, M gene, F gene, HN gene and L gene, wherein each of the M gene, the F gene and the FIN gene is from a Sendai virus strain Cl.151-derived gene and wherein at least one of the M gene, the F gene and the HN gene is functionally deleted and the L gene encodes the amino-acid sequence of the L protein in which the amino-acid residue at position 1618 is valine and a method of producing the same.

Abstract: Stem cell reprogramming genes cloned into a single sustained expression-type Sendai viral vector are shown to reprogram differentiated somatic cells into induced pluripotent stem (iPS) cells without integration of vector sequences into the host cell's genome. The genes are transduced into normal differentiated somatic cells via infection with recombinant Sendai virus. After expression of the reprogramming genes and subsequent induction of pluripotency, the vector genome RNA including the reprogramming genes is removed from the cell to establish an iPS cell that is genetically identical to the parent somatic differentiated cell thus reducing the risk of tumorigenic transformation caused by random integration of vector sequences into the host genome. The method promises to provide safe, autologous iPS cells that can be used for human cell replacement and regeneration therapeutic applications.

Abstract: A persistently infective virus vector is produced by using a gene so modified as to encode an amino acid sequence including a valine substituted for an amino acid residue at position-1618 in the amino acid sequence for an L protein of a persistently non-infective Sendai virus. A non-transmissible, persistently infective virus vector is also produced by defecting or deleting at least one of M gene, F gene, and HN gene. These virus vectors have no cytotoxicity, can achieve the sustained gene expression over a long period of time, is safe, and is therefore useful.

Abstract: A substance of interest is contained in nanospheres which are then encapsulated in fusogenic liposomes to prepare transport carriers that allow physiologically active substances, especially those having high molecular weight such as proteins and genes, to be introduced into cells efficiently and which permit the introduced active substance to be released in the cell at controlled rate. The fusogenic liposomes are prepared by conferring the fusogenic capability of Sendai virus to known liposomes.

Abstract: A &lgr; phage with a nuclear localization signal has been obtained by constructing a vector capable of expressing a fused protein between a gpD protein constituting the head of a &lgr; phage and a nuclear localization signal sequence, transforming Escherichia coli with this vector, and propagating a mutant &lgr; phage which cannot express the gpD protein in E. coli in this transformant. It has been confirmed that the resulting &lgr; phage is capable of packaging &lgr; phage DNAs of 80% and 100% genome sizes. After further confirming that the nuclear localization signal exposed on the outside of the head of this phage, this phage has been microinjected into cells to analyze its nuclear localization activity. Thus, it has been clarified that this phage has a nuclear localization activity.

Abstract: The present invention provides a novel phage expressing in its head a bi-functional protein that has nuclear translocation and cell adhesion activities. The phage is used to package a foreign substance such as a gene. As a bi-functional protein, TAT protein of HIV can be used. The phage is useful in gene therapy.

Abstract: A &lgr; phage with a nuclear localization signal has been obtained by constructing a vector capable of expressing a fused protein between a gpD protein constituting the head of a &lgr; phage and a nuclear localization signal sequence, transforming Escherichia coli with this vector, and propagating a mutant &lgr; phage which cannot express the gpD protein in E. coli in this transformant. It has been confirmed that the resulting &lgr; phage is capable of packaging &lgr; phage DNAs of 80% and 100% genome sizes. After further confirming that the nuclear localization signal exposed on the outside of the head of this phage, this phage has been microinjected into cells to analyze its nuclear localization activity. Thus, it has been clarified that this phage has a nuclear localization activity.

Abstract: A &lgr; phage with a nuclear localization signal has been obtained by constructing a vector capable of expressing a fused protein between a gpD protein constituting the head of a &lgr; phage and a nuclear localization signal sequence, transforming Escherichia coli with this vector, and propagating a mutant &lgr; phage which cannot express the gpD protein in E. coli in this transformant. It has been confirmed that the resulting &lgr; phage is capable of packaging &lgr; phage DNAs of 80% and 100% genome sizes. After further confirming that the nuclear localization signal exposed on the outside of the head of this phage, this phage has been microinjected into cells to analyze its nuclear localization activity. Thus, it has been clarified that this phage has a nuclear localization activity.

Abstract: A &lgr; phage with a nuclear localization signal has been obtained by constructing a vector capable of expressing a fused protein between a gpD protein constituting the head of a &lgr; phage and a nuclear localization signal sequence, transforming Escherichia coli with this vector, and propagating a mutant &lgr; phage which cannot express the gpD protein in E. coli in this transformant. It has been confirmed that the resulting &lgr; phage is capable of packaging &lgr; phage DNAs of 80% and 100% genome sizes. After further confirming that the nuclear localization signal exposed on the outside of the head of this phage, this phage has been microinjected into cells to analyze its nuclear localization activity. Thus, it has been clarified that this phage has a nuclear localization activity.