Engineered U7 Small Nuclear RNA Restores Correct β-Globin Pre-mRNA Splicing in Mouse βIVS2-654-Thalassemic Erythroid Progenitor Cells

Hum Gene Ther. 2021 May;32(9-10):473-480. doi: 10.1089/hum.2020.145. Epub 2020 Nov 2.

Abstract

Restoration of correct splicing of βIVS2-654-globin pre-mRNA was previously accomplished in erythroid cells from β-thalassemia/HbE patients by an engineered U7 small nuclear RNA (snRNA) that carried a sequence targeted to the cryptic branch point and an exonic splicing enhancer, U7.BP+623 snRNA. In this study, this approach was tested in thalassemic mice carrying the βIVS2-654 mutation. While correction of βIVS2-654 pre-mRNA splicing was achieved in erythroid progenitors transduced with a lentiviral vector carrying the U7.BP+623 snRNA, a high level of truncated U7.BP+623 snRNA was also observed. The discrepancy of processing of the modified U7 snRNA in human and mouse constructs hamper the evaluation of pathologic improvement in mouse model.

Keywords: RNA splicing; U7 snRNA; thalassemia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Erythroid Precursor Cells / metabolism
  • Humans
  • Mice
  • RNA Precursors* / genetics
  • RNA Precursors* / metabolism
  • RNA Splicing / genetics
  • RNA, Small Nuclear / genetics
  • RNA, Small Nuclear / metabolism
  • beta-Globins* / genetics

Substances

  • RNA Precursors
  • RNA, Small Nuclear
  • U7 small nuclear RNA
  • beta-Globins