Abstract: A tool for the isolation of c-kit expressing cells is provided. This tool consists of a c-kit plasmid targeting vector which is capable of integrating into the wild type c-kit allele and encodes a chimeric fluorescent protein comprising a nucleolar localization signal such as, TCOF-12, RLP313, RPS254 or Fxr2h5. The construct generates a condensed, bright, fluorescent signal that can be localized in living tissue and after dissociation. The construct allows visualization using confocal microscopy and allows automated cell sorting of the dissociated cells using amongst others flow cytometry.

Abstract: The present invention provides methods for inducing insulin gene expression in cultured pancreas cells, the method comprising contacting a culture of endocrine pancreas cells expressing a PDX-1 gene with a GLP-1 receptor agonist, wherein the cells have been cultured under conditions such that the cells are in contact with other cells in the culture, thereby inducing insulin gene expression in the ceils. The invention also provides methods of treating a diabetic human subject using the methods of the invention.

Abstract: The invention relates to isolated human pluripotent adult stem cells which express CD13, CD34, CD56 and CD117, and which do not express CD1O, which are capable of differentiating in all three germ lineages and differentiated cells derived therefrom.

Abstract: The present invention describes a novel recombinant NADH recycling system that is used as a process for producing reduced compounds. In a specific embodiment, the reduced compounds include ethanol, succinate, lactate, a vitamin, a pharmaceutical and a biodegraded organic molecule. The NADH recycling system effects metabolic flux of reductive pathways in aerobic and anaerobic environments.

Abstract: The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising of the steps of: compartmentalizing genetic elements into microcapsules; expressing the genetic elements to produce their respective gene products within the microcapsules; sorting the genetic elements which produce the gene product having a desired activity. The invention enables the in vitro evolution of nucleic acids by repeated mutagenesis and iterative applications of the method of the invention.

Abstract: A method of screening cellular polypeptides for pro-apoptotic or anti-apoptotic activity in a cell of a particular cell-type by: (a) culturing cells of the particular cell-type under non apoptotic conditions and culturing cells of the particular cell-type under apoptotic conditions, and (b) determining subcellular localization of the cellular polypeptides in the cultured cells, wherein a localization of a cellular polypeptide in lipid rafts in cultured cells under non apoptotic conditions and a segregation of the cellular polypeptide from lipid rafts in cultured cells under apoptotic conditions is indicative that the cellular polypeptide has a pro-apoptotic or an anti-apoptotic activity in the particular cell-type.

Abstract: An artificial promoter library (or a set of promoter sequences) for a selected organism or group of organisms is constructed as a mixture of double stranded DNA fragments, the sense strands of which comprise at least two consensus sequences of efficient promoters from said organism or group of organisms, or parts thereof comprising at least half of each, and surrounding intermediate nucleotide sequences (spacers) of variable length in which at least 7 nucleotides are selected randomly among the nucleobases A, T, C and G. The sense strands of the double stranded DNA fragments may also include a regulatory DNA sequence imparting a specific regulatory feature, such as activation by a change in the growth conditions, to the promoters of the library. Further, they may have a sequence comprising one or more recognition sites for restriction endonucleases added to one or both of their ends.

Abstract: The invention provides for cells containing nucleic acids which include lysozyme gene expression controlling region nucleotide sequences which typically are linked to a polynucleotide encoding a heterologous polypeptide.

Abstract: The invention provides for lysozyme gene expression control regions which may include a 5? matrix attachment region; an intrinsically curved region of DNA; a transcription enhancer; a negative regulatory element; at least one hormone responsive element; an avian CRI repeat element; a proximal lysozyme promoter, and can be linked to a nucleotide sequence encoding a heterologous polypeptide.

Abstract: The present invention relates to codon optimised polynucleotides which are efficiently expressed in mammalian cells and encode insect proteins from Dermaphagoids dust mite. In particular, the optimised codon polynucleotides encode a protein from Dermaphagoides pteronyssinus, such as DerP1 or proDerP1. The present invention also provides methods of preparing pharmaceutical compositions comprising the expression of the codon optimised polynucleotides, and vectors and transformed host cells comprising them.

Abstract: The present invention provides methods and compositions for drug screens to identify and characterize agents that are agonistic or antagonistic to activation of the promoter region of the NAG-1 gene. Activation of the NAG-1 gene is associated with the apoptotic elimination of cancer cells both in vitro and in vivo. The invention also provides novel promoter region sequences of the NAG-1 gene.

Abstract: The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising of the steps of: (a) compartmentalising genetic elements into microcapsules; (b) expressing the genetic elements to produce their respective gene products within the microcapsules; (c) sorting the genetic elements which produce the gene product having a desired activity.

Abstract: The invention concerns the introduction of predetermined genetic changes in target genes of a living cell by introducing an oligodeoxynucleotide encoding the predetermined change. The oligodeoxynucleotides are effective in animal, plant and bacterial cells. Specific end modifications that greatly increase the effectiveness of the oligodeoxynucleotides in bacteria are described. Surprisingly, unmodified oligodeoxynucleotides can be as effective in mammalian cells, including in vivo hepatocytes, as the modified nucleotides and can be as effective or more effective than chimeric oligonucleotides that consist of a mixture of deoxynucleotides and 2?-O-methyl ribonucleotides.

Abstract: 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, a 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.

Abstract: The present invention is a composition and process for avoiding non-specific uptake of targeted liposomal complexes in the lung and other highly vascular issues. The reversible masking of liposomal complexes allows increased delivery of nucleic acid molecules to target cells or tissues.

Abstract: Disclosed herein are methods for detecting multiple compounds in a sample, involving: (a) contacting the sample with a mixture of binding reagents, the binding reagents being nucleic acid-protein fusions, each having (i) a protein portion which is known to specifically bind to one of the compounds and (ii) a nucleic acid portion which encodes the protein portion and which includes a unique identification tag; (b) allowing the protein portions of the binding reagents and the compounds to form complexes; (c) capturing the binding reagent-compound complexes; (d) amplifying the nucleic acid portions of the complexed binding reagents; and (e) detecting the unique identification tag of each of the amplified nucleic acids, thereby detecting the corresponding compounds in the sample. Also disclosed herein are kits for carrying out such methods.