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Evaluering af Exon integration induceret af splejse skifte Antisense oligonukleotider i SMA patienten fibroblaster
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Médecine
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Journal JoVE Médecine
Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts

Evaluering af Exon integration induceret af splejse skifte Antisense oligonukleotider i SMA patienten fibroblaster

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

Chapitres

  • 00:05Titre
  • 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

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Traduction automatique

Forskellige antisense oligonukleotider (AONs) har vist sig at fremkalde exon integration (splice modulation) og redde SMN udtryk for spinal muskelatrofi (SMA). Her, beskriver vi en protokol for AON lipotransfection til at fremkalde exon integration i SMN2 gen og evalueringsmetoder for at fastlægge effekten i SMA patient fibroblaster.

Tags

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

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