Screening method of target sequence capable of inhibiting NFAT gene and application of screening method

Abstract

The invention belongs to the technical field of gene engineering, and discloses a screening method of a target sequence capable of inhibiting NFAT gene and an application of the screening method. Thescreening method of the target sequence capable of inhibiting NFAT gene comprises the following steps of designing a genetic sequence to screen shRNA optimal target sequence; constructing a plasmid vector and a reverse transcription virus vector; transfecting prostate cancer cells; and screening out a target sequence with the optimal inhibition effect on the NFAT gene. A negative control of base mismatch and a negative control of a chaos sequence are set, so that shRNA can be fully proved to only generate specific gene silencing on a target gene, and the accuracy of screening of the target sequence capable of inhibiting the NFAT gene is enhanced; by transfecting prostate cancer cells, the target sequence with the inhibition effect on the NFAT gene can be obtained, the screening process issimpler and more convenient, and the obtained target cell has a better inhibition effect on the NFAT gene.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a screening method for a target sequence capable of inhibiting NFAT gene and its application. Background technique [0002] At present: Nuclear factor of activated T cells (NFAT) is a family of transcription factors that play an important role in inducing gene transcription in immune responses. In addition to T cells, this type of protein can also be expressed on many immune cells, such as B lymphocytes, mast cells, eosinophils, etc. Its activity is regulated by calcium-dependent calmodulin phosphatase C. NFAT is a transcription factor with multidirectional regulatory functions, such as regulating the activation, differentiation and self-tolerance of T cells. NFAT is expressed in a variety of immune cells, such as T cells, B cells, NK cells, mast cells, monocytes and eosinophils, etc., and induces the transcription of cytokines and other genes in the bod...

AI Technical Summary

Problems solved by technology

However, the current screening methods for inhibiting NFAT genes have poor accuracy and cannot accurately obtain target sequences that have an inhibitory effect on NFAT genes, and the screening products have poor inhibitory effects on NFAT genes

[0003] Through the above analysis, the existing problems and defects of the prior art are: the accuracy of the current screening method for inhibiting NFAT genes is poor, and it is impossible to accurately obtain the target sequence that has an inhibitory effect on the NFAT gene, and the inhibitory effect of the screened product on the NFAT gene Difference

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