Isolation tool for viable c-kit expressing cells

Details Isolation tool for viable c-kit expressing cells Isolation tool for viable c-kit expressing cells

C - Chemistry, Metallurgy

12

N

C12N 15/85 (2006.01) A01K 67/027 (2006.01) C12N 5/10 (2006.01) C12N 15/09 (2006.01) C12N 15/79 (2006.01) C12N 15/90 (2006.01) C12Q 1/02 (2006.01) C12Q 1/04 (2006.01) C12Q 1/68 (2006.01) G01N 21/64 (2006.01)

Patent

CA 2468778

The present invention provides a tool for the isolation of ck-it expressing cells. 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 localisation signal such as, TCOF- 12, RLP313, RPS254 or Fxr2h5.. Said construct generates a condensed, bright, fluorescent signal that can be localised in living tissue and after dissociation. The construct allows visualisation using confocal microscopy and allows automated cell sorting of the dissociated cells using amongst others flow cytometry.

L'invention concerne un outil permettant l'isolation de cellules d'expression ck-it. Cet outil comprend un vecteur de ciblage plasmide c-kit qui est capable d'intégration dans le type sauvage c-kit allele et codant pour une protéine fluorescente chimère comprenant un signal de localisation nucléolaire tel que TCOF-1¿2?, RLP31¿3?, RPS25¿4? ou Fxr2h¿5.?. Ladite construction génère un signal fluorescent brillant condensé qui peut être localisé dans des tissus vivants et après dissociation. La construction permet la visualisation par microscopie confocale et permet le triage automatisé de cellules pour les séparer des cellules dissociées, en particulier par cytométrie de flux.

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