This invention relates to the field of therapeutics. Most specifically invention provides methods of generating in vitro engineered immune cells conditionally expressing interleukin-12 (IL-12) and one or more immunomodulators under the control of a gene expression modulation system in the presence of activating ligand and uses for therapeutic purposes in animals.

The present invention relates to nucleic acid constructs comprising selectable marker genes in a multicistronic transcription unit for use in the generation and selection of eukaryotic host cells for expression of a gene product of interest. For increased stringency of selection, the coding sequence of the selectable marker may be directed preceded by a relatively short functional open reading frame to reduce the efficiency of translation of the selectable marker, and/or the amino acid sequence of the selectable marker may comprise one or more mutations that reduce the level of resistance provide by the mutated marker as compared to its wild type counterpart. The invention further relates to methods for generating eukaryotic host cells for expression of a gene product of interest, wherein these nucleic acid constructs are used, and to methods for producing a gene product of interest wherein thus generated host cells are applied.

There are provided a recombinant cell that enables efficient production of protein, a method for producing the same, and a method for producing protein by using the cell. The cell is a mammalian cell having integrated therein 3 or more expression units containing an exogenous gene of interest, wherein gap elements are provided among multiple expression units, wherein the gap elements are designed to have chain lengths of 0.5 kbp or more, and the gap elements are mutually different sequences. Such a cell can produce recombinant protein stably for a long period and, especially, can show a strong positive linear correlation between the number of genes of interest introduced therein and the expression levels.

This invention relates to the field of therapeutics. Most specifically invention provides methods of generating in vitro engineered immune cells conditionally expressing interleukin-12 (IL-12) and one or more immunomodulators under the control of a gene expression modulation system in the presence of activating ligand and uses for therapeutic purposes in animals.

The disclosure provides methods and compositions useful for obtaining induced stem cells, methods of making and use thereof.

Provided are nucleic acid constructs and systems which comprise (i) a first nucleic acid construct encoding a toxin operatively linked to a first promoter and at least one cancer-associated signaling responsive enhancer element; and (ii) a second nucleic acid construct encoding an anti-toxin operatively linked to a second promoter, the second promoter being stronger than the first promoter.

Also provided are pharmaceutical compositions comprising same and methods of using same for treating cancer.

The invention relates to an infectious arenavirus particle that is engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells. One or more of the four arenavirus open reading frames glycoprotein (GP), nucleoprotein (NP), matrix protein Z and RNA-dependent RNA polymerase L are removed or mutated to prevent replication in normal cells but still allowing gene expression in arenavirus vector-infected cells, and foreign genes coding for an antigen or other protein of interest or nucleic acids modulating host gene expression are expressed under control of the arenavirus promoters, internal ribosome entry sites or under control of regulatory elements that can be read by the viral RNA-dependent RNA polymerase, cellular RNA polymerase I, RNA polymerase II or RNA polymerase III. The modified arenaviruses are useful as vaccines and therapeutic agents for a variety of diseases.

A method of treating cancer, a viral infection and/or an immune system disorder, comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a vector, wherein the vector comprises (a) nucleic acid sequences encoding 4-1BB ligand (4-1BBL), single chain IL-12 (scIL-12) and IL-2, and (b) at least one regulatory nucleic acid sequence providing for an increased expression level of 4-1BBL as compared to the expression levels of scIL-12 and IL-2, and other related methods.

The disclosure provides methods and compositions useful for obtaining induced stem cells, methods of making and use thereof.

Methods are provided for the ex vivo reprogramming of adult mammalian cells, wherein the genes used in reprogramming the adult cells are expressed with heterologous promoters that increase expression of associated genes while the cell is in a fetal or adult non-regenerative state, but down-regulated the expression of genes once cells reach a regenerative state and before the cells are reprogrammed to pluripotency. In addition, heterologous promoters uniquely expressing genes when cells are in an embryonic (pre-fetal state) are used to increase expression of toxic gene products in cancer cells.