Methods and drugs for targeted editing of RNA based on leaper technology

Abstract

The present application relates to a nucleic acid drug based on LEAPER technology, and a method for using the nucleic acid drug to target editing RNA to treat diseases such as Hurler syndrome. The method includes using the nucleic acid drug to edit the base from adenosine to hypoxanthine on the RNA, accurately repairing the pathogenic G>A mutation site such as Hurler syndrome, and restoring The normal expression of the encoded protein such as IDUA in vivo.

Description

technical field [0001] The present application belongs to the field of gene editing therapy, and in particular, relates to a method for editing RNA based on LEAPER (Leveraging Endogenous ADAR for Programmable Editing on RNA) technology and repairing disease-causing mutations, which includes using LEAPER technology to perform on RNA from A to Site-directed editing of I bases to prevent or treat diseases caused by G>A mutations such as Herler's syndrome. Background technique [0002] Hurler syndrome (Hurler syndrome), also known as mucopolysaccharide storage disease IH type (Mucopolysaccharides IH, MPS IH), is the most serious of the three subtypes IH, IH / S, and IS of MPSI type. It is an autosomal recessive genetic disease (autosomal recessive genetic disease), which is caused by the deficiency of α-L-iduronidase (IDUA) in the body of the patient recessive, AR). The root cause of Heller's syndrome is the mutation of the IDUA gene encoding the IDUA protein located at 4p16....

AI Technical Summary

Problems solved by technology

First, exogenously expressed nucleases are often required to have a large molecular weight, which drastically reduces the efficiency of their delivery into the body by viral vectors

Secondly, due to the exogenous expression of nucleases, there is a potential nuclease off-target possibility, which will make its application a potential carcinogenic risk

Finally, exogenously expressed nucleases are found in bacteria and not naturally occurring in humans or mammals, which makes it possible to elicit an immune response in the patient, which may cause damage to the patient itself, or It may also neutralize exogenously expressed nucleases, thereby losing their proper activity or hindering further therapeutic interventions

[0006] In 2017, Zhang Feng's research group reported an RNA editing technology called REPAIR (RNA Editing for Programmable Ato I Replacement) (RNA editing with CRISPR-Cas13, Cox et al., 2017), which expresses Cas13 exogenously -ADAR fusion protein and a single guide RNA (single guide RNA, sgRNA) can also achieve the editing from A to I of the target RNA, but this method, like CRISPR technology, still requires the expression of foreign proteins

The complementary RNA used in this method is short (less than 54nt), but requires complex chemical modifications, and the editing efficiency is not high

However, the RESTORE technology requires the presence of IFN-γ to have high editing efficiency, and IFN-γ is a key factor determining the development and severity of autoimmunity (Interferon-γ and systemic autoimmunity., Pollard et al., 2013, This makes the application of the technology in the medical field greatly discounted

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