Position dependent recognition of gnn nucleotide triplets by...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/12 (2006.01) C07K 14/47 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2429555

The specificity of binding of a zinc finger to a triplet or quadruplet nucleotide target subsite depends upon the location of the zinc finger in a multifinger protein and, hence, upon the location of its target subsite within a larger target sequence. The present disclosure provides zinc finger amino acid sequences for recognition of triplet target subsites having the nucleotide G in the 5'-most position of the subsite, that have been optimized with respect to the location of the subsite within the target site. Accordingly, the disclosure provides finger position-specific amino acid sequences for the recognition of GNN target subsites. This allows the construction of multi-finger zinc finger proteins with improved affinity and specificity for their target sequences, as well as enhanced biological activity.

La spécificité de liaison d'un doigt de zinc à un sous-site cible de triplet ou de quadruplet nucléotidique dépend de l'emplacement du doigt de zinc dans une protéine à doigts multiples et, par conséquent, de l'emplacement du sous-site cible dans une séquence cible plus grande. L'invention concerne des séquences d'acides aminés de doigts de zinc servant à reconnaître les sous-sites cibles de triplets qui présentent le nucléotide G en position 5' d'extrémité du sous-site, ces séquences ayant été optimisées quant à l'emplacement du sous-site dans le site cible. L'invention concerne aussi des séquences d'acides aminés spécifiques de position de doigt destinées à reconnaître des sous-sites cibles GNN ; et permet de construire des protéines à multiples doigts de zinc qui présentent une affinité et une spécificité accrues pour leurs séquences cibles, ainsi qu'une activité biologique accrue.

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