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Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts
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Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts

Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts

DOI:
10.3791/57530-v

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07:02 min

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May 11, 2018

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, ,
1Department of Medical Genetics,University of Alberta Faculty of Medicine and Dentistry, 2Muscular Dystrophy Canada Research Chair, Department of Medical Genetics,University of Alberta Faculty of Medicine and Dentistry

Chapters

  • 00:05Title
  • 01:00LNA/DNA Mixmer Transfection Followed by One-step RT-PCR to Measure the Exon Inclusion Rate of SMN2
  • 05:07Results: Use of LNA/DNA Mixmers to Study Efficient Inclusion of SMN2 Exon and Rescue of SMN Protein Expression
  • 06:40Conclusion

Summary

Automatic Translation

Automatic Translation

Various antisense oligonucleotides (AONs) have been shown to induce exon inclusion (splice modulation) and rescue SMN expression for spinal muscular atrophy (SMA). Here, we describe a protocol for AON lipotransfection to induce exon inclusion in the SMN2 gene and the evaluation methods to determine the efficacy in SMA patient fibroblasts.

Tags

Keywords: Exon InclusionSplice SwitchingAntisense OligonucleotidesSMAFibroblastsLipotransfectionTransfectionLNA/DNA MixmerSerum-deprived MediaTransfection ReagentCell ConfluencyGPC ReagentRNA Extraction

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