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Method for carrying out protein translation by using circular RNA and application of circular RNA

A circular and reaction system technology, applied in the field of protein translation and its application, can solve problems such as the construction of restricted expression systems for gene therapy applications

Active Publication Date: 2020-09-29
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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Problems solved by technology

A common method is to use viral IRES to initiate the translation of circular RNA. However, due to the possible immune rejection of pathogenic viral RNA in the host, and the RNA elements derived from viruses basically contain complex RNA secondary structures and long sequences. , which limits the construction of expression system based on viral IRES and the later application of gene therapy

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  • Method for carrying out protein translation by using circular RNA and application of circular RNA
  • Method for carrying out protein translation by using circular RNA and application of circular RNA
  • Method for carrying out protein translation by using circular RNA and application of circular RNA

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preparation example Construction

[0139] The invention provides a method for in vitro protein synthesis, comprising the steps of:

[0140] (i) providing the reaction system described in the second aspect of the present invention;

[0141] (ii) Incubate the synthesis system in step (i) for a period of time T1 under suitable conditions, thereby synthesizing the protein.

[0142] In another preferred example, the method further includes: (iii) optionally isolating or detecting the protein from the reaction system.

[0143] The main advantages of the present invention include:

[0144](1) The present invention firstly develops a set of methods for designing and synthesizing a new type of artificially synthesized eukaryotic translation initiation element of non-viral origin with high translation activity and at the same time sequence structure and length controllable, and uses this to translate the initiation element Drives translation of circular RNAs.

[0145] (2) The present invention screens for the first ti...

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Abstract

The invention provides a method for carrying out protein translation by using circular RNA and an application of the circular RNA. Particularly, the invention provides a circular RNA construct. The circular RNA construct has a structure as shown in a formula I of TI-Z1-Z2 (I) in the 5'-3' direction; in the formula, TI is a translation initiation element, Z1 is an expression cassette expressing foreign protein, and Z2 is none or other element. The circular RNA construct can significantly enhance the translation efficiency in vivo and in vitro.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to protein translation using circular RNA and its application. Background technique [0002] Protein is the most important biomacromolecule in organisms, and its mutation or abnormal expression can lead to diseases. Therefore, corresponding diseases can be treated through protein replacement or expression. Common protein replacement or expression therapies include delivery systems based on ribonucleic acid (DNA) vectors, delivery systems based on deoxyribonucleic acid (RNA) vectors, and protein delivery systems. These methods all require the translation of messenger RNA to produce protein. The common translation initiation method in eukaryotes is cap-dependent translation, which initiates translation mainly through the translation initiation factor recognizing the special cap structure at the 5′ end of messenger RNA, and this type of translation only exists in linear messenger RNA . ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/63C12N5/10C12N1/19
CPCC12N15/11C12N15/63C12N15/67C12N2840/203C12P21/02C12N15/111
Inventor 王泽峰杨赟樊晓娟
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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