Artificially modified AKT3 circular RNA and application thereof in new crown vaccine

A circular, artificial technology, applied in the direction of DNA/RNA vaccination, whole cell/virus/DNA/RNA components, positive-sense single-stranded RNA virus, etc., can solve the problem of low processing efficiency, RNA from genomic sites related to human diseases problems, such as easy storage, not easy to degrade by RNase, and high translation efficiency

Active Publication Date: 2021-04-30
KUNMING MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite inefficient processing and mostly expressed at low levels, some circular RNAs originate from genomic loci associated with human disease, and accumulating evidence points to their potential roles in transcriptional, post-transcriptional and translational regulation

Method used

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  • Artificially modified AKT3 circular RNA and application thereof in new crown vaccine

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Experimental program
Comparison scheme
Effect test

Embodiment

[0012] 1. Synthesis of DNA template fragments

[0013] First, through the whole genome synthesis technology, the synthetic sequence is the DNA template shown in SEQ ID NO.3, which includes the artificially modified AKT3 circular RNA sequence and the sequence of the spike protein of the new coronavirus.

[0014] 2. PCR amplification of DNA template fragments

[0015] Perform PCR amplification with primers whose sequences are SEQ ID NO.4 and SEQ ID NO.5 to obtain a large number of double-stranded DNA templates. Note that high-fidelity enzymes are used for amplification, and PCR products are purified after amplification.

[0016] 3. RNA synthesis based on template fragments

[0017] The T7 RNA Polymerase of the T7 promoter is used for amplification, and the template is the PCR product in the previous step. After the amplification is completed, the DNA is digested with RNase-free DNase, and then the obtained RNA is purified.

[0018] 4. Circularization of linear RNA to obtain ci...

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Abstract

The invention discloses an artificially modified AKT3 circular RNA (ribonucleic acid) and application thereof in a new crown vaccine, which are characterized in that the modified AKT3 circular RNA has a transcription starting function, a section of protein open reading frame is added in the middle, so that the circular RNA can stably express the protein of the protein open reading frame, and if a corresponding immunogen protein open reading frame is added, the protein of the protein open reading frame can be stably expressed. The monoclonal antibody can act on a carrier of a circRNA vaccine, so that an immune animal generates an antibody, and the monoclonal antibody can be used as a vaccine for prevention or treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine and biotechnology, and in particular relates to an artificially modified AKT3 circular RNA, which can express the protein after the insertion site of the circular RNA is inserted into an open reading frame, and can be used as a carrier of various vaccines. Background technique [0002] The structure and properties of RNA are very unstable, which makes more and more people pay attention to its therapeutic use. Although the molecule is extremely sensitive to the nearly ubiquitous ribonucleases (RNases), mRNA was first promoted as a therapeutic agent in 1989 following the development of widely used in vitro transfection techniques. Only a few years later, mRNA was promoted as a vaccine platform for therapeutics. Combining the immunological features of live attenuated vaccines (such as endogenous antigen expression and T cell induction) with those of inactivated or subunit vaccines (such as defined composi...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12N15/85C12N15/50A61K39/215A61P31/14
CPCC12N15/85C07K14/005A61P31/14C12N2800/107C12N2770/20022C12N2770/20034A61K2039/53
Inventor何越峰孙明军毛一竹蒋成兰谭婧文周倩尹锦瑶
OwnerKUNMING MEDICAL UNIVERSITY