Construction method of monoclonal cell line with specific gene knockout of porcine small intestine cells
The invention belongs to the technical field of gene engineering, and discloses a construction method and application of a monoclonal cell line with a specific gene knocked out of porcine small intestine cells, firstly, a gene editing vector is constructed, and gene editing can be carried out aiming...
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Main Authors | , , , , , , , |
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Format | Patent |
Language | Chinese English |
Published |
14.05.2024
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Subjects | |
Online Access | Get full text |
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Summary: | The invention belongs to the technical field of gene engineering, and discloses a construction method and application of a monoclonal cell line with a specific gene knocked out of porcine small intestine cells, firstly, a gene editing vector is constructed, and gene editing can be carried out aiming at specific sites of genes; the rAAV-SaCas9-eGFP-sgRNA virus with strong small intestine tropism is packaged, so that the gene editing efficiency of cells is improved; and screening to obtain the specific gene knockout cell model of the porcine small intestine cells through rapid identification of eGFP fluorescence by flow cytometry. And a single cell strain is obtained by a cloning method, so that the monoclonal cell line with the specific gene knocked out of the porcine small intestine cells is obtained. According to the rAAV-SaCas9-eGFP-sgRNA virus disclosed by the invention, porcine small intestine cells are infected, and a method for rapidly establishing a monoclonal cell line with a specific gene knocked out |
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Bibliography: | Application Number: CN202410117232 |