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TEDx Perth talk on Morpholinos and Duchenne muscular dystrophy (Wilton, late 2016)

Prof. Steve Wilton: TEDx Perth talk on Morpholinos and Duchenne muscular dystrophy.

If you have had time with Steve, life is better. He's a good sort.

Thursday, February 16, 2017 - 01:12
Which splice junction to target, predicting cryptic splice sites, and doing the two-non-overlapping oligo specificity experiment

I've had the pleasure of corresponding with Yuhong Liang (梁雨虹)of 4A Biotech in China; she is the product manager for Morpholino oligos at 4A Biotech. We have been working with 4A for a little over a year now and Yuhong has been learning very quickly, but is still new enough to the Morpholino field that she is asking questions of broad interest. She had some questions regarding which splice junction to target, predicting cryptic splice sites, and the process of doing the two-non-overlapping oligo specificity experiment.

Wednesday, February 8, 2017 - 01:07
Getting Morpholinos into cultured cells

There are three usual options to consider for cell culture experiments with Morpholinos. There are other methods, but most labs choose one of these.

(1) If your cells can tolerate electroporation, that is an inexpensive option for delivery of standard Morpholino oligos. However, electroporation causes high cell mortality for many cell types.

Friday, December 16, 2016 - 04:44
Validating Morpholino phenotypes with CRISPRs

I've been enjoying a conversation with Martin Blum about Morpholinos, CRISPR mutants, funding and publication. He wrote:

Good morning Jon,

My study section at the German Research Council DFG now accepts MO-proposals, no problem. Still, many people particularly working in zebrafish are too careless with controls and rescues. That might backfire at one point in the future.

My take is that we will use CRISPR to validate MOs and then use MOs most of the time.

Best wishes,
Martin

Thursday, December 8, 2016 - 04:37
Assessing activity of a splice-modifying Morpholino

This is about detecting the activity of a splice-modifying oligo by reverse-transcriptase PCR and gel electrophoresis.

The expected outcome of an exon targeting an exon 3 splice junction (either i2e3 or e3i3) is elimination of exon 3 from the mature mRNA (along with both of its flanking introns, i2 and i3). The typical way to assess activity of a splice-modifying oligo is to isolate RNA from treated samples and run reverse-transcriptase PCR (RT-PCR), then run the PCR product on a gel to assess the mass of the product against a DNA ladder.

Friday, December 2, 2016 - 03:49
Nippon Shinyaku Morpholino Treatment for DMD: FDA fast track

FDA Grants Fast Track Designation to NS-065/NCNP-01 for the Treatment of Duchenne Muscular Dystrophy.

http://www.nippon-shinyaku.co.jp/english/company_profile/news.php?id=3113

Tuesday, November 22, 2016 - 06:51
Translational development of splice-modifying antisense oligomers - Review

Translational development of splice-modifying antisense oligomers.
Fletcher S, Bellgard MI, Price L, Akkari AP, Wilton SD.
Expert Opin Biol Ther. 2016 Nov 2:1-16. [Epub ahead of print]
http://www.tandfonline.com/doi/full/10.1080/14712598.2017.1250880

Friday, November 4, 2016 - 07:20
(review) Loss-of-function genetic tools for animal models: cross-species and cross-platform differences

Housden BE, Muhar M, Gemberling M, Gersbach CA, Stainier DY, Seydoux G, Mohr SE, Zuber J, Perrimon N. Loss-of-function genetic tools for animal models: cross-species and cross-platform differences. Nat Rev Genet. 2016 Oct 31. doi: 10.1038/nrg.2016.118. [Epub ahead of print]

Wednesday, November 2, 2016 - 04:29
Acute RNAi versus knockout: mutation triggers compensation

No Morpholino work in this one, but it explores a difference between targeting RNA versus DNA.

Cell-Intrinsic Adaptation Arising from Chronic Ablation of a Key Rho GTPase Regulator.
Cerikan B, Shaheen R, Colo GP, Gläßer C, Hata S, Knobeloch KP, Alkuraya FS, Fässler R, Schiebel E.
Dev Cell. 2016 Sep 28. pii: S1534-5807(16)30595-0. doi: 10.1016/j.devcel.2016.08.020.

http://www.sciencedirect.com/science/article/pii/S1534580716305950

Tuesday, October 4, 2016 - 11:35
Morpholino drug approved by FDA

The US Food and Drug Administration today (19 Sep 2016) granted Accelerated Approval to eteplirsen (EXONDYS 51), a Morpholino oligo-based treatment for some forms of Duchenne muscular dystrophy (DMD). This is the first approval of a Morpholino drug.

Monday, September 19, 2016 - 15:31

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