%0 Journal Article
%T Restoration of correct 汕IVS2-654-globin mRNA splicing and HbA production by engineered U7 snRNA in 汕-thalassaemia/HbE erythroid cells
%J -
%D 2019
%R https://doi.org/10.1038/s41598-019-43964-3
%X A cytosine to thymine mutation at nucleotide 654 of human 汕-globin intron 2 (汕IVS2-654) is one of the most common mutations causing 汕-thalassaemia in Chinese and Southeast Asians. This mutation results in aberrant 汕-globin pre-mRNA splicing and prevents synthesis of 汕-globin protein. Splicing correction using synthetic splice-switching oligonucleotides (SSOs) has been shown to restore expression of the 汕-globin protein, but to maintain therapeutically relevant levels of 汕-globin it would require lifelong administration. Here, we demonstrate long-term splicing correction using U7 snRNA lentiviral vectors engineered to target several pre-mRNA splicing elements on the 汕IVS2-654-globin pre-mRNA such as cryptic 3∩ splice site, aberrant 5∩ splice site, cryptic branch point and an exonic splicing enhancer. A double-target engineered U7 snRNAs targeted to the cryptic branch point and an exonic splicing enhancer, U7.BPˋ+ˋ623, was the most effective in a model cell line, HeLa IVS2-654. Moreover, the therapeutic potential of the vector was demonstrated in erythroid progenitor cells derived from 汕IVS2-654-thalassaemia/HbE patients, which showed restoration of correctly spliced 汕-globin mRNA and led to haemoglobin A synthesis, and consequently improved thalassaemic erythroid cell pathology. These results demonstrate proof of concept of using the engineered U7 snRNA lentiviral vector for treatment of 汕-thalassaemia
%U https://www.nature.com/articles/s41598-019-43964-3