Targeted SMN Exon Skipping: A Useful Control to Assess In Vitro and In Vivo Splice-Switching Studies

Biomedicines

Sue Fletcher¹  LorenL. Flynn¹  IantheL. Pitout¹  SteveD. Wilton¹  Abbie Adams¹  Anja Stirnweiss²  Chalermchai Mitrpant³ 

[1] Centre for Molecular Medicine and Innovative Therapeutics, Health Futures Institute, Murdoch University, Murdoch, WA 6150, Australia;PYC Therapeutics, Nedlands, WA 6009, Australia;Perron Institute for Neurological and Translational Science, Nedlands, WA 6009, Australia;

关键词: antisense oligonucleotide;    morpholino;    positive control;    survival motor neuron;    cell penetrating peptide;   

DOI  :  10.3390/biomedicines9050552

来源: DOAJ

【 摘 要 】

The literature surrounding the use of antisense oligonucleotides continues to grow, with new disease and mechanistic applications constantly evolving. Furthermore, the discovery and advancement of novel chemistries continues to improve antisense delivery, stability and effectiveness. For each new application, a rational sequence design is recommended for each oligomer, as is chemistry and delivery optimization. To confirm oligomer delivery and antisense activity, a positive control AO sequence with well characterized target-specific effects is recommended. Here, we describe splice-switching antisense oligomer sequences targeting the ubiquitously expressed human and mouse SMN and Smn genes for use as control AOs for this purpose. We report two AO sequences that induce targeted skipping of SMN1/SMN2 exon 7 and two sequences targeting the Smn gene, that induce skipping of exon 5 and exon 7. These antisense sequences proved effective in inducing alternative splicing in both in vitro and in vivo models and are therefore broadly applicable as controls. Not surprisingly, we discovered a number of differences in efficiency of exon removal between the two species, further highlighting the differences in splice regulation between species.

【 授权许可】

Unknown