Nonhuman model animal unresponsive to immunopotentiating...

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/00 (2006.01) A01K 67/027 (2006.01) A61P 37/04 (2006.01) C07K 14/705 (2006.01) C12N 15/85 (2006.01) G01N 33/50 (2006.01)

Patent

CA 2468011

It is intended to provide a nonhuman model animal unresponsive to a synthetic compound which lacks a gene function encoding TLR7 recognizing an immunopotentiating synthetic compound such as imidazoquinoline on its chromosome. A gene fragment of the whole gene sites including the intracellular domain and the transmembrane domain or a part thereof of a TLR7 gene obtained from a mouse gene library is substituted by a plasmid having polyA signal and a marker gene to construct a targeting vector. Then this targeting vector is linearized and transferred into embryo stem cells. The target embryo stem cells lacking the TLR7 gene function are microinjected into a mouse blastocyst to construct a chimeric mouse. Then this chimeric mouse is crossed with a wild type mouse to give a heterozygote mouse. Next, the heterozygote mouse is intercrossed and thus a TLR7 knockout mouse is obtained.

L'invention concerne un modèle animal non humain ne réagissant pas à un composé synthétique auquel il manque une fonction génique codant TLR7, permettant de reconnaître un composé synthétique d'immunopotentialisation, notamment de l'imidazoquinoline sur son chromosome. Un fragment génique des sites géniques entiers comprend le domaine intracellulaire et le domaine transmembranaire ou une partie de ceux-ci d'un gène TLR7 obtenu à partir d'une banque de gènes de souris, est substitué par un plasmide présentant un signal polyA et un gène marqueur pour construire un vecteur ciblant. Puis ce vecteur ciblant est linéarisé et transféré dans des cellules souches embryonnaires. Ces cellules souches embryonnaires cibles auxquelles il manque la fonction génique TLR7 sont micro-injectées dans un blastocyste de souris pour construire une souris chimérique. Cette souris chimérique est croisée avec une souris de type sauvage pour donner une souris hétérozygote. Puis, cette souris hétérozygote est intercroisée, et une souris k.o. est obtenue.

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