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Outcome: Targeted exon is skipped or not Question answered:Can the oligo induce exon skipping? Summary: Use of oligonucleotides to change splicing Tool.

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Presentation on theme: "Outcome: Targeted exon is skipped or not Question answered:Can the oligo induce exon skipping? Summary: Use of oligonucleotides to change splicing Tool."— Presentation transcript:

1 Outcome: Targeted exon is skipped or not Question answered:Can the oligo induce exon skipping? Summary: Use of oligonucleotides to change splicing Tool to shift alternative splice patterns, correct aberrant splicing, reframe transcripts and knockdown genes Overview: oligos to induce exon skipping Normal splicing Intended splicing Step 1: oligo design Targets: splice sites, branch point sequence or exon internal sequences A Step 2: Transfect oligo into relevant cultured cells A oligo Step 3: Isolate RNA Step 4: RT-PCR analysis Step 5: Sequence analysis

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3 MNT12345678 910C Skip Full length B. Confirm skip by sequence analysis Exon 27Exon 29 C.Lab-Chip analysis MM Full length 50 nmol/L Skip 125 nmol/L MM Full length 300 nmol/L Skip 80 nmol/L Skip % = 80 300+80 = 21% Skip % = 125 125+50 = 71% 12345678910 D. Compare AONs A. Agarose analysis


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