Method for enrichment and/or isolation of dopaminergic neurons

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 5/10 (2006.01) C12N 15/09 (2006.01) C12Q 1/02 (2006.01) C12Q 1/04 (2006.01) C12Q 1/68 (2006.01) C12N 5/06 (2006.01)

Patent

CA 2381065

The invention of this application provides a method comprising introducing a reporter nucleic acid molecule that expresses a fluorescent protein under control of the promoter/ enhancer of a gene that is expressed in dopaminergic neurons, into each of cells, and isolating fluorescence-emitting cells. The invention also provides a method for visualizing and identifying dopaminergic neurons alive that exist with in cells, which comprises introducing the above-mentioned reporter nucleic acid molecule into each of cells, and measuring the fluorescence distribution within the cells. The invention further provides a method for identifying a dopaminergic neurons-inducing factor, which comprises introducing the reporter nucleic acid molecule into cells that have the ability to differentiate into dopaminergic neurons, then incubating the cells with a candidate substance, and determining whether the candidate substance is a dopaminergic neurons-inducing factor by using the fluorescence of the cells as an indicator.

L'invention concerne un procédé qui consiste à transférer des molécules d'acide nucléique exprimant une protéine fluorescente sous l'effet de la régulation par le promoteur/amplificateur d'un gène exprimé dans les neurones dopaminergiques, dans les cellules individuelles d'une masse cellulaire, et à séparer les cellules fluorescentes de ladite masse cellulaire. L'invention porte également sur un procédé qui consiste à transférer les molécules d'acide nucléique susmentionnées dans des cellules individuelles d'une masse cellulaire, pour permettre la mesure de la répartition de la fluorescence de ladite masse, visualiser et identifier ainsi les neurones dopaminergiques présents dans la masse cellulaire à l'état viable. L'invention se rapporte également à un procédé d'identification d'un facteur inducteur de neurone dopaminergique, qui consiste à transférer une molécule d'acide nucléique dans des cellules capables de se différentier en neurones dopaminergiques, ce qui permet auxdites cellules de co-exister avec une substance candidate, puis à déterminer si ladite substance candidate est un facteur inducteur de neurone ou non, la fluorescence des cellules étant utilisée comme un indication.

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