Reca / rad51 recombinase-mediated production of recombinant...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/90 (2006.01) A01K 67/027 (2006.01) C12N 5/06 (2006.01) C12N 9/10 (2006.01) C12N 15/10 (2006.01) C12N 15/82 (2006.01) C12N 15/87 (2006.01)

Patent

CA 2399366

A method comprising: a) altering a chromosomal sequence of a donor nucleus of a donor cell by introducing a pair of single-stranded targeting polynucleotides, and a recombinase into said donor nucleus of said donor cell, wherein said pair of targeting polynucleotides are substantially complementary to each other and each comprising a homology clamp that substantially corresponds to or is substantially complementary to a predetermined DNA sequence of said nucleus; and, b) transplanting said nucleus into an oocyte to produce a recombinant zygote. A method of altering a nucleic acid sequence of a mitochondria or chloroplast of a cell comprising: introducing into a cell a pair of single-stranded targeting polynucleotides, and recombinase, wherein said pair of targeting polynucleotides are substantially complementary to each other, and each comprising a homology clamp that substantially corresponds to or is substantially complementary to a predetermined nucleic acid sequence of said mitochondria or chloroplast, whereby said sequence is altered.

La présente invention concerne un procédé consistant à a) modifier une séquence chromosomique d'un noyau donneur d'une cellule donneuse en introduisant une paire de polynucléotides de ciblage à simple brin et une recombinase dans ledit noyau donneur de ladite cellule donneuse, les polynucléotides de la paire de polynucléotides de ciblage étant sensiblement complémentaires l'un à l'autre et comprenant chacun un crochet d'homologie qui correspond sensiblement/est sensiblement complémentaire à une séquence d'ADN dudit noyau; et, b) transplanter ledit noyau dans un ovocyte pour produire un zygote recombiné. En outre, cette invention concerne un procédé de modification de la séquence d'acide nucléique d'une mitochondrie ou d'un chloroplaste d'une cellule, consistant à: introduire dans une cellule une paire de polynucléotides de ciblage à simple brin et une recombinase, les polynucléotides de la paire de polynucléotides de ciblage étant sensiblement complémentaires l'un à l'autre, et comprennent chacun un crochet d'homologie qui correspond sensiblement/est sensiblement complémentaire à la séquence d'acide nucléique prédéterminée des mitochondrie ou chloroplaste précités, ladite séquence étant modifiée.

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