| 摘要 |
The invention provides methods and compositions for generating a homozygous mutation at genomic loci of polyploid cells, e.g., mammalian cells. The methods of the invention employ a gene search vectors comprising a selection marker linked to and under the control of a regulated promoter. A gene search vector is inserted into a genomic locus of a cell to produce a single allelic mutation. Double allelic mutation at the genomic locus is achieved by generating and selecting cells that have undergone homologous recombination that leads to homozygous insertion of the gene search vector, or portion thereof, at the genomic locus by culturing the polyploid cells under a concentration of an inducing agent to which the regulated promoter is responsive such that the activity level of the regulated promoter is tuned, e.g., reduced from full strength, to facilitate selection of cells containing homozygous mutation at the locus. |
