C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/55 (2006.01) C12N 9/78 (2006.01) C12Q 1/68 (2006.01) G01N 33/50 (2006.01)
Patent
CA 2580011
The invention relates to a method of accelerating the induction of somatic mutations in vitro. The inventive method comprises the expression of at least one cDNA expressing a modified version of the AID gene in the cells to be mutated, in culture conditions and a medium that are suited thereto, said modified version resulting from an AID gene in which the three hydrophobic amino acids, leul89, phe193 and leul96, have been replaced by means of alanine mutation in each case. The invention can be used to induce mutations in Burkitt's lymphoma BL2. The invention can also be used to induce mutations in the immunoglobulin genes of immortalised antibody-producing cells, such as mouse hybridoma cells, human hybridoma cells or human B-cell lines immortalised by the Epstein-Barr virus (EBV).
L'invention concerne un procédé pour l'accélération de l'induction de mutations somatiques in vitro, comportant l'expression dans les cellules à faire muter, dans des conditions de culture et dans un milieu appropriés aux dites cellules, d'au moins un ADNc exprimant une version modifiée du gène AID, ladite version modifiée étant faite d'un gène AID dans lequel on a remplacé les trois acides aminés hydrophobes leul89, phe193 et leul96 par mutation en alanine dans chaque cas. Application à l'induction de mutations sur le lymphome de Burkitt BL2. Application à l'induction de mutations dans les gènes d'immunoglobulines de cellules immortalisées productrices d'anticorps, notamment les hybridomes de souris, les hybridomes humains ou les lignées B humaines immortalisées par le virus Epstein-Barr (EBV).
Reynaud Claude-Agnes
Weill Jean-Claude
Goudreau Gage Dubuc
Institut Necker
Reynaud Claude-Agnes
Weill Jean-Claude
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