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|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.
|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.
|Tuesday, November 22, 2016 - 06:51|
|Translational development of splice-modifying antisense oligomers - Review||
Translational development of splice-modifying antisense oligomers.
|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.
|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|
|miRNA targeting and second oligo specificity controls||
Once again, a conversation with a new Morpholino user led to a discussion others might find useful.
For the two non-overlapping oligo specificity control, you do two separate sets of injections of the two oligos targeting the same miRNA. If the embryos treated with the oligo targeting the 5' end of the miRNA produces the same phenotype as the oligo targeting the 3' end of the miRNA, then that is good: it supports the idea that the phenotype you are seeing is caused by knocking down the activity of the miRNA you intend to target, and not caused by binding to an unexpected RNA.
|Friday, September 9, 2016 - 09:59|
|Vivo-Morpholino use in rodent livers: annotated citations||
In response to a question about the history of Vivo-Morpholino use in rodent livers, I assembled this annotated citation list.
|Wednesday, August 31, 2016 - 10:47|
|phase 1 clinical trial with a Morpholino||
Here's a report of a phase 1 clinical trial with a Morpholino oligo.
Komaki H, Nagata T, Saito T, Masuda S, Takeshita E, Tachimori H, Sasaki M, Takeda S. Exon 53 skipping of the dystrophin gene in patients with Duchenne muscular dystrophy by systemic administration of NS-065/NCNP-01: A phase 1, dose escalation, first-in-human study. Neuromuscular Disord. 2015;26(2)S261-2. doi:10.1016/j.nmd.2015.06.276
|Tuesday, July 19, 2016 - 13:57|