A method for evolving a cell having desired phenotype and...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/04 (2006.01) C12N 15/10 (2006.01) C12N 15/66 (2006.01) C12Q 1/02 (2006.01)

Patent

CA 2474146

The present invention relates to evolution of a cell or a composition of cells having a desired property or functionally. The principle behind the evolution of cells according to the invention is to produce a great diversity of genes in each cell subjected to evolution and a great diversity of genes among the cells in a composition according to the invention and to exchange the genes between the cells from time to time. In preferred embodiments the genes are arranged in expression cassettes in concatemers in the cells, as well as in artificial chromosomes. Said methods comprising the steps of a) obtaining a composition of cells, at least one cell of said composition comprising a1) at least two expressible nucleotide sequences, at least one of said sequences being incorporated into an artificial chromosome in the cell and/or a2) at least two expression cassettes of the following formula: [rs2-SP-PR-X-TR-SP- rs11 wherein rsl and rs2 together denotes a restriction site, SP individually denotes a spacer, PR denotes a promoter, capable of functionning in the first cell, X denotes an expressible nucleotide sequence, TR denotes a terminator, and/or a3) at least two expressible nucleotide sequences, said sequences being heterologous to the cell, determining at least one screening functionality. The genes are preferably co-ordinately controllable for increasing diversity. The desired property of functionality is preferably a compound, or a series of compounds synergistically acting to each other.

La présente invention concerne la mise au point d'une cellule ou d'une composition de cellules ayant une propriété ou une fonctionnalité souhaitée. La mise au point de cellules décrite dans cette invention permet, d'une part, de produire une grande diversité de gènes dans chaque cellule soumise à cette évolution et une grande diversité de gènes parmi les cellules contenues dans une composition décrite dans l'invention et, d'autre part, d'échanger ces gènes entre des cellules, le cas échéant. Dans des modes de réalisation privilégiés, les gènes sont placés dans des cassettes d'expression en concatémères dans des cellules et dans des chromosomes artificiels. Les gènes sont, de préférence, régulés de manière coordonnée afin d'en augmenter la diversité. La propriété ou la fonctionnalité souhaitée est, de préférence, un composé, ou une série de composés qui agissent de manière synergétique les uns sur les autres.

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