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Application of codon-optimized trichosanthin gene in gene therapy

A trichosanthin, codon optimization technology, applied in gene therapy, application, genetic engineering and other directions, to achieve the effect of improving the level of transgene expression and the effect of large biological effects

Inactive Publication Date: 2015-11-04
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of a codon-optimized Trichosanthin gene (coTCS) in gene therapy has not been reported yet.

Method used

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  • Application of codon-optimized trichosanthin gene in gene therapy
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  • Application of codon-optimized trichosanthin gene in gene therapy

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Embodiment 1

[0038] 1. Experimental materials and methods

[0039] 1. Plasmids, primers

[0040] The pdsAAV-AFPp-EGFP plasmid was constructed by Glushakova et al. (Mingozzi F, High KA: Therapeutic in vivo gene transfer for genetic disease using AAV: progress and challenges. Nature reviews Genetics 2011, 12(5):341-355.). Among them, AFPp is the alpha-fetoprotein promoter, and EGFP is the enhanced green fluorescent protein. The TCS gene was synthesized by Life Technologies, based on the published sequence (http: / / www.ncbi.nlm.nih.gov / nuccore / M34858.1; T.kirilowiitrichosanthin (TCS) mRNA, complete cds; GenBank: M34858.1) , And use restriction enzymes Age I and Hind III for subcloning, and put them into pdsAAV-AFPp-EGFP, pdsAAV-CBAp-EGFP plasmids. Among them, CBAp is the chicken β-actin promoter. All plasmids were sequenced before use. coTCS is a variety of solutions to obtain TCS gene optimization through software provided by Integrated DNA Technologies and Life Technologies. In the solutions,...

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Abstract

The invention relates to a codon-optimized trichosanthin gene and application thereof in gene therapy. According to the invention, after codon optimization, the trichosanthin gene is subjected to drug delivery through a viral or non-viral vector, can express protein at a higher level in mammalian cells so as to play a greater biological role. In the invention, the drug delivery carrier is optimized to improve its transgenic expression level in mammalian cells and play a maximum antitumor role, thus having important significance to research and promotion of tumor gene therapy.

Description

Technical field [0001] The present invention relates to the field of gene therapy, specifically, a codon-optimized trichosanthin gene and its application in gene therapy. Background technique [0002] Trichosanthin (TCS) is derived from the traditional Chinese medicine Radices trichosanthis. Trichosanthes is the root of Trichosanthes kirilowii Maxim or Trichosanthes rosthornii Herms. The effects of Trichosanthes are: clearing away heat and purging fire, producing body fluids and quenching thirst, reducing swelling and draining pus, and mainly treating: quenching thirst, jaundice, fever and thirst, dryness and hemoptysis. TCS is composed of 247 amino acid residues. It belongs to type I Ribosome-inactivating proteins (RIP) with a molecular weight of about 27kDa. It has many functions such as inhibiting the growth of a variety of tumor cells, antiviral, immune regulation, and anti-AIDS virus. Kind of pharmacological activity. Clinically, TCS has been used for labor induction and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/63C12N15/861A61K48/00A61P35/00
Inventor 凌昌全张院辉王园凌晨殷惠霞
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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