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Short hairpin rna, recombinant vector and application for nfat3 gene target

A technology of recombinant vectors and short hairpins, which is applied in the direction of DNA/RNA fragments, retroRNA viruses, applications, etc., can solve the problems of short-term inhibition of target gene expression, low transfection efficiency, and non-permanent expression of siRNA, to achieve inhibition proliferating effect

Inactive Publication Date: 2020-09-22
SHENZHEN UNIV
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Problems solved by technology

[0005] Initially, RNA interference used the method of synthesizing siRNA in vitro, but there were disadvantages such as low transfection efficiency, inability to express siRNA transferred into cells persistently, and short-term inhibition of target gene expression, which limited its application.

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  • Short hairpin rna, recombinant vector and application for nfat3 gene target
  • Short hairpin rna, recombinant vector and application for nfat3 gene target
  • Short hairpin rna, recombinant vector and application for nfat3 gene target

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Embodiment Construction

[0026] The present invention will be further described below through specific embodiments in conjunction with the accompanying drawings.

[0027] The methods used in this embodiment are conventional methods unless otherwise specified, and the specific steps can be found in "Molecular Cloning: A Laboratory Manual" (Sambrook, J., Russell, David W., 3rdedition, 2001, NY, Cold Spring Harbor) . All DNA primers used were synthesized by Invitrogen; melanoma cells were purchased from ATCC and preserved by the applicant; various restriction enzymes and Taq enzymes were purchased from TaKaRa Company; T4 DNA ligase was purchased from NEB Company; gel recovery kit and plasmid The extraction kit was purchased from OMEGA Company; DMEM medium and fetal bovine serum were purchased from HyClone Company; the transfection reagent Lipofectamine was purchased from Invitrogen Company; the melanoma cell culture medium was DMEM medium containing 10% fetal bovine serum. Unless otherwise specified, ot...

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Abstract

The invention discloses shRNA for an NFAT3 gene target, a recombinant vector and an application. A sequence of the gene target is shown in SEQ ID NO:3; a positive-strand sequence of RNA is shown in SEQ ID NO:1; and a negative-strand sequence of RNA is shown in SEQ ID NO:2. The shRNA sequence is designed for the NFAT3 gene target, and the corresponding recombinant vector is constructed; the recombinant vector can perform transcription in host cells to obtain NFAT3-shRNA, siRNA can be generated after processing of the host cells, and mRNA of NFAT3 is targeted, so that NFAT3 expression is reduced, a gene silencing effect is caused, and the proliferation of melanoma cells is effectively inhibited; and the recombinant vector has an anti-cancer application value, and provides a foundation for further research of functions of NFAT3 in skin cancer cells and NFAT3-targeted drug research and development.

Description

technical field [0001] The invention relates to the fields of molecular biology, genetic engineering technology and biomedicine, in particular to a short hairpin RNA targeting the NAT3 gene target, a recombinant vector and its application. Background technique [0002] Skin cancer is one of the most common clinical malignant tumors, which can be roughly divided into malignant melanoma and non-melanoma skin cancer, the latter mainly including squamous cell carcinoma and basal cell carcinoma. Severe cases of skin cancer can be life-threatening, and its incidence is increasing year by year, which has become a global public health problem. Multiple risk factors can contribute to the development of skin cancer, including endogenous factors (genetic factors, gene mutations, etc.) and exogenous factors (such as ultraviolet exposure, chemical carcinogens, and other environmental stressors). Early diagnosis and treatment can reduce the death and deformity caused by the disease. How...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/867A61K48/00A61P35/00
CPCA61K48/0058C12N15/113C12N15/86C12N2310/141C12N2310/531C12N2740/15043
Inventor 郑多肖田黄诗莹胡跃明
Owner SHENZHEN UNIV
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