THE FROG PRINCE, A TRANSPOSON VECTOR FOR GENE TRANSFER IN VERTEBRATES

• Main Authors: IVICS, ZOLTAN, IZSVAK, ZSUZSANNA, MISKEY, CSABA, PLASTERK, RONALD
• Format: Patent
• Language: English; French; German
• Published: 16.07.2008
• Subjects: AGRICULTURE; ANIMAL HUSBANDRY; BEER; BIOCHEMISTRY; CARE OF BIRDS, FISHES, INSECTS; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FISHING; FORESTRY; HUMAN NECESSITIES; HUNTING; HYGIENE; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; NEW BREEDS OF ANIMALS; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; SPIRITS; TRAPPING; VINEGAR; WINE

The present invention relates to a transposon-based DNA integration system comprising (a) a transposon which is devoid of a polynucleotide encoding a functional transposase and which comprises a polynucleotide of interest, wherein the transposon comprises inverted repeats having a degree of identity with the repeats within SEQ ID NO: 2 and its inverted repeat, respectively, of at least 90%; and (b) a transposase having at its N-terminus a DNA binding domain comprising the sequences of SEQ ID NO: 3 and 4; or (c) a polynucleotide encoding the transposase of (b). The present invention further relates to a method of transferring a polynucleotide of interest into cells of a vertebrate comprising the step of introducing the transposon-based DNA integration system of the invention into said cells. In addition, the invention relates to a method of effecting RNAi comprising (a) stably introducing a transposon comprising an expression cassette expressing a short interfering RNA and a selectable marker gene as part of the transposon-based DNA integration system of the invention into a cell; (b) selecting for cells expressing the selectable marker; and (c) assessing whether the transcription/translation of the desired gene is effected by RNAi. A further embodiment of the invention is a method of gene trapping genes comprising the steps of (a) introducing the transposon-based DNA integration system of the invention into a cell; and (b) assessing for the expression of a selectable marker wherein expression of a selectable marker is indicative of integration of the transposon into a transcibed gene of the cell.