Modified human u1snrna molecule, a gene encoding for the modified human u1snrna molecule, an expression vector including the gene, and the use thereof in gene therapy of familial dysautonomia and spinal muscular atrophy

Abstract

The invention provides a modified human U1snRNA molecule, capable of correcting the skipping of an exon caused by a mutation localized in the sequence comprised between 50 base pairs upstream and 20 base pairs downstream of an exon, wherein a portion of a single-stranded nucleotide sequence of the 5′ region of the wild-type human U1snRNA is replaced by a single-stranded binding nucleotide sequence, wherein the binding nucleotide sequence is selected from the group consisting of: uggcgcuua, aauggcgcu, aguacaauggcgc (SEQ ID NO: 87), gcaaacaguacaau (SEQ ID NO: 88), ucgcaaacaguaca (SEQ ID NO: 89), gcaaacagu, cuagucgcaaac (SEQ ID NO: 90), uacaaaaguaagauuca (SEQ ID NO: 83), aaaccauaaaguuuuacaa (SEQ ID NO: 84) and caaaccauaaaguuuua (SEQ ID NO: 96).

Description

FIELD OF THE INVENTION[0001]The present invention concerns modified human snRNA molecules (hereinafter designated as Exon Specific U1—ExSpeU1), which are suitable to be used in gene therapy methods. In particular, the invention relates to snRNA molecules capable of correcting aberrant splicing processes caused by genetic mutations and related to human diseases with different case histories, which are often very serious.BACKGROUND OF THE INVENTION[0002]Many human genetic diseases (about 15%) are caused by genetic mutations that, by interfering with the correct messenger RNA intracellular maturation, compromise the accurate subsequent protein biosynthesis and induce synthesis of non-functional proteins. Mostly, the point mutations accountable for splicing defects concern gene sequences that are critical for the recognition of the primary transcript by the machinery appointed for processing the same. The donor and acceptor sites located at the exon-intron boundaries, as well as gene-sp...

AI Technical Summary

Benefits of technology

The invention is about a type of RNA molecule called U1snRNA, which can selectively target and act on specific parts of a gene called exons. These exons are more unique and different from the parts that connect them called introns. Surprisingly, the U1snRNA molecules can still induce the inclusion of exons even when there are mutations in the gene. This can help researchers better understand the function of these genes and potentially develop new treatments for genetic diseases.

Problems solved by technology

As a consequence of CFTR modification, the transfer of salts through cell membranes is compromised, mainly causing a production of secretions that could be defined as “dehydrated”: a sweat very rich in sodium and chlorine and a dense and viscous mucus that tends to obstruct the ducts, compromising the function of various organs and systems.

To date, no treatment for this disease, which generally leads to death at an age that depends on the seriousness of the case history, has yet been identified.

However, expression of SMN2 is impaired by a synonymous mutation in the exon which results in an aberrant maturation of the messenger RNA with consequent skipping of exon 7 and inactivation of the gene itself.

However, in neurons, this amount is not sufficient to support a physiological activity, leading to the pathological condition.

However, this method presents a certain degree of non-specificity of action of the therapeutic snRNA molecule towards the target gene, due to the relative conservation of the 5′ss sites and consequent risk of interfering with the maturation of transcripts generated from other functional wild-type genes.

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