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MAMMALIAN CELL LINE FOR PROTEIN PRODUCTION AND LIBRARY GENERATION
MAMMALIAN CELL LINE FOR PROTEIN PRODUCTION AND LIBRARY GENERATION
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机译:用于蛋白质生产和图书馆生成的哺乳动物细胞系
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摘要
According to a first aspect of the invention, a method for the generation of a cell line is provided, comprising the steps of (a) providing a plurality of mammalian B cells, wherein each of the plurality of B cells comprises a transgenic genomic DNA sequence encoding a marker protein inserted into an endogenous immunoglobulin locus comprised in said B cell, and wherein the transgenic genomic DNA sequence is amenable to cleavage by a site directed nuclease, particularly Cas9; (b) replacing the transgenic genomic DNA sequence encoding a marker protein with a second transgenic DNA sequence encoding a protein of interest; (c) sorting B cells based on the presence or absence of the marker protein; and (d) collecting B cells in which the marker protein is absent.
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