Method for preparing circular RNA and nucleic acid sequence for method

By inserting tag sequences into both end arms of the linear RNA precursor and using affinity chromatography media for separation, the problem of efficient preparation and purification of circular RNA in the existing technology has been solved, and an efficient, economical and safe process has been achieved. Circular RNA production.

WO2025011502A1PCT designated stage expired Publication Date: 2025-01-16RINUAGENE BIOTECHNOLOGY CO LTD +1
View PDF 7 Cites 0 Cited by

Patent Information

Application Number
PCT/CN2024/104101
Authority / Receiving Office
WO · WO
Patent Type
Applications
Current Assignee / Owner
Priority Date
2023-07-07
Filing Date
2024-07-05
Publication Date
2025-01-16

AI Technical Summary

Technical Problem

It is difficult to efficiently prepare high-purity circular RNA in the prior art, especially on the scale of industrial production, and conventional purification methods have problems such as low loading, high cost, complex process and low resolution.

Method used

By inserting tag sequences into both end arms of the linear RNA precursor, the specific binding of the tag sequence to the affinity chromatography medium is used to achieve the separation of circular RNA and by-products. The specific method includes generating linear RNA precursors with poly(A) tags, performing self-splicing and cyclization reactions, followed by contact with affinity chromatography media, separating and collecting the supernatant to obtain high-purity circular RNA.

Benefits of technology

This method can significantly improve the production efficiency, purity and yield of circular RNA, reduce production costs, simplify the process, improve the safety of circular RNA, basically eliminate linear RNA by-products, and meet the needs of industrial production.

✦ Generated by Eureka AI based on patent content.

Smart Images

  • Figure CN2024104101_16012025_PF_FP_ABST
    Figure CN2024104101_16012025_PF_FP_ABST
Patent Text Reader

Abstract

Provided is a method for preparing a circular RNA, which method comprises inserting purification tags, such as poly(A) tags or functional variants thereof into two terminal arms of a linear RNA precursor, and separating the circular RNA, which is obtained by means of the cyclization reaction of the linear RNA precursor, from other byproducts of the cyclization reaction as impurities using an affinity chromatographic medium capable of simultaneously binding to the tags. Further provided are a linear RNA precursor with a purification tag inserted in a specific region and an encoding DNA thereof. The insertion of the purification tag in the specific region does not affect the yield of linear RNA precursors produced by means of the in-vitro transcription (IVT) of the encoding DNA.
Need to check novelty before this filing date? Find Prior Art

Citation Information

Patent Citations

  • Method for efficiently purifying and identifying cyclic RNA based on second-generation Illumina sequencing platform

    CN110283895A

  • Vector for preparing circular RNA (Ribonucleic Acid) and application thereof

    CN114507691A

  • Cyclic RNA compositions and methods

    CN115052635A

  • Cyclic RNA compositions and methods

    CN116034114A

  • Cyclic RNA compositions and methods

    CN116113419A