Therapeutic potential of antisense oligonucleotides as modulators of alternative splicing

Abstract

Correction of splicing by antisense oligonucleotides β-Globin. β-Thalassemia is a serious genetic blood disorder (3) in which production of β-globin, a subunit of hemoglobin, is partially or entirely ablated by mutations in the β-globin gene. The defect decreases the oxygen-carrying capacity of red blood cells and causes compensatory expansion of the bone marrow An estimated 60% of all human genes undergo alternative splicing, a highly regulated process that produces splice variants with different functions. Such variants have been linked to a variety of cancers, and genetic diseases such as thalassemia and cystic fibrosis. This Perspective describes a promising approach to RNA repair based on the use of antisense oligonucleotides to modulate alternative splicing and engender the production of therapeutic gene products.

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“…Our work relies on 2'O-methyl RNA oligonucleotides as splice-site ASOs, and joins a growing list of synthetic nucleic acid-based compounds that have been used to modify splicing [40]. ASOs employing a combination of phosphorothioate and 2'O-methyl RNA modifications have been used to modify dystrophin splicing [11,12], ß-globin splicing [14], and SMN2 splicing [41].…”

Alternative splicing and disease

“…Special chemistries were devised to prevent RNAseH-mediated cleavage of the RNA and to lower toxicity (reviewed in [149, 150]). A major drawback of the therapeutic use of oligonucleotides is their delivery and uptake in the cells.…”

Direct Intraventricular Delivery of Drugs to the Rodent Central Nervous System

“…This specific method takes advantage of the mouse ventricular system that bathes the entire brain and spinal cord, allowing widespread distribution of the ASOs. We use ASOs that are 18-20 mer RNA-like molecules that directly bind the target mRNA sequence and, depending on the ASO chemical modifications, either A) recruit RNase-H to degrade the mRNA leading to knockdown or B) shift alternative splicing 1,2,16,17 . It should be noted that multiple molecules exist for knocking down a specific protein in vivo, including shRNA.…”