Method for transfecting pig T lymphocytes in vitro by applying nuclear transfection method

Abstract

The invention relates to a method for transfecting pig T lymphocytes in vitro by applying a nuclear transfection method, comprising the following steps of: introducing an expression vector containing foreign genes or mRNA (Messenger RNA) of the foreign genes into the pig T lymphocytes by applying an electroporation technology and a cell-specific nuclear transfection solution; culturing transfected cells, and separating the pig T lymphocytes expressing the foreign genes. The method disclosed by the invention has the advantages of high transfection efficiency, easiness and convenience for operation, less needed plasmid quantity and fast expression of target genes and is beneficial to researching the specific gene functions of T cells at activated and effective stages.

Description

technical field [0001] The invention relates to a method for in vitro transfection of porcine T lymphocytes, in particular to a method for in vitro transfection of porcine T lymphocytes using a nuclear transfection method. Background technique [0002] Transgenic technology is an important means to study the gene function of mammalian cells, including retrovirus-mediated infection, liposome-mediated transfection, microinjection, electrotransfection, etc. T cells are cells that are difficult to transfect, and it is difficult for exogenous nucleic acid substances to be expressed in T cells. After decades of hard work, people have established a relatively complete and effective method for transfecting T cells. These methods mainly include transfection and electroporation mediated by virus particles such as lentivirus and adenovirus. However, these methods have their inherent defects. First, most of these methods are developed based on human and mouse T cells, and are not full...

AI Technical Summary

Problems solved by technology

However, these methods have their inherent defects. First of all, most of these methods are developed based on human and mouse T cells, and are not fully applicable to porcine T lymphocytes.

Secondly, these methods need to pre-stimulate T cells before transfection, and cannot be used to effectively study the specific gene function of T cell activation phase

Thirdly, the virus-mediated transfection method has high efficiency, but packaging the virus requires high-tech and high-cost support, which is not conducive to promotion

At the same time, taking into account the current status of China's pig industry, that is, my country is a large pork production and consumption country, and the current epidemics such as swine fever, reproductive and respiratory syndrome (PRRS) are still seriously prevalent, and multi-pathogen cross-infection often occurs , the disease resistance of pigs and the prevention and control effect of vaccines are significantly reduced. It is necessary to develop a new transgenic method suitable for transfection of pig-derived T cells, and to study the relevant immune functions of pig T cells.

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