Transgenic mice expressing fluorescent protein under the...

A - Human Necessities – 01 – K

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A01K 67/00 (2006.01) A01K 67/027 (2006.01) C07K 14/47 (2006.01) C12N 5/00 (2006.01) C12N 5/10 (2006.01) C12N 15/10 (2006.01) C12N 15/63 (2006.01) C12N 15/85 (2006.01)

Patent

CA 2392051

Non-human transgenic mammals are produced which have, incorporated in their genome, DNA which includes a regulatory sequence of a mammalian nestin gene, operably linked to a gene coding for a marker/reporter protein. The regulatory sequence can include a promoter and a sequence present in the second intron of the mammmalian nestin gene. Preferably, the marker/reporter protein is a fluorescent protein, for example a green fluorescent protein, modified for enhanced fluorescence. Multipotent and, in particular, neural stem and progenitor cell populations are observed in the organs of the non-transgenic mammal or progeny thereof. Multipotent stem and progenitor cells are isolated directly from the non-human transgenic mammal, progeny or embryo thereof, for example by FACS, without culture passages.

L'invention concerne la production de mammifères transgéniques non humains qui possèdent, incorporé dans leur génome, un ADN comprenant une séquence régulatrice du gène nestine d'un mammifère relié fonctionnement à un gène codant un protéine marqueur/rapporteur. Cette séquence régulatrice peut renfermer un promoteur et une séquence présente dans le second du gène nestine d'un mammifère. De préférence, la protéine marqueur/rapporteur est une protéine fluorescente, notamment une protéine fluorescente verte, que l'on a modifiée pour qu'elle présente une fluorescence améliorée. On observe des populations de cellules multipotentes, notamment des populations de cellules souches neuronales ou de progéniteurs, dans les organes du mammifère non transgénique ou de sa progéniture. Les cellules souches multipotentes et les progéniteurs sont isolés directement du mammifère transgénique humain, de sa progéniture ou d'un embryon dudit mammifère, par exemple par la technique FACS (fluorescent activated cell sorting), sans passage par une phase de culture.

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