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(paper): Deletion of morpholino binding sites (DeMOBS) to assess specificity of morphant phenotypes

Another specificity control strategy involving mutants and Morpholinos

Cunningham CM, Bellipanni G, Habas R, Balciunas D. Deletion of morpholino binding sites (DeMOBS) to assess specificity of morphant phenotypes. Sci Rep. 2020 Sep 21;10(1):15366. doi: 10.1038/s41598-020-71708-1.
https://www.nature.com/articles/s41598-020-71708-1

Paper with links to resources for designing splice-switching oligos

Marchalot A, Lambert J-M, Boyer F, Pollet J, Moreau J, Feuillard J, Faumont N, Delpy L. Splice switching oligonucleotide mediated gene knockdown in B cells and plasma cells. bioRxiv. 2020;[preprint] doi:10.1101/2020.09.18.302984

https://www.biorxiv.org/content/10.1101/2020.09.18.302984v1

Links to resources for designing splice-switching oligos

Specificity test failed: Morpholino in a mutant

This paper presents an example of a Morpholino off-target effect shown by testing the Morpholino in a CRISPR mutants.

Jiang Z, Carlantoni C, Allanki S, Ebersberger I, Stainier DYR. Tek/Tie2 is not required for cardiovascular development in zebrafish. Development. 2020 Sep 14:dev.193029. doi: 10.1242/dev.193029. Online ahead of print.

https://dev.biologists.org/content/early/2020/09/12/dev.193029

Targeting alternative splicing to upregulate protein expression

Potential application in antisense therapeutics: upregulation of targeted protein expression

Here is a new paper demonstrating techniques for targeting steric-blocking antisense oligos to alternatively-spliced transcripts undergoing NMD to alter the splicing and recover transcripts encoding useful proteins. This is a technique for antisense upregulation of a protein activity.

Lim KH, Han Z, Jeon HY, Kach J, Jing E, Weyn-Vanhentenryck S, Downs M, Corrionero A, Oh R, Scharner J, Venkatesh A, Ji S, Liau G, Ticho B, Nash H, Aznarez I. Antisense oligonucleotide modulation of non-productive alternative splicing upregulates gene expression. Nat Commun. 2020;11(1):3501. doi:10.1038/s41467-020-17093-9

https://www.nature.com/articles/s41467-020-17093-9

Retention of the last intron

Here are a few publications addressing Morpholinos and retention of the last intron.

See the section: Retention of wnt11b Intron 4 Recapitulates the Failure to Form Somites in tra2b Morphants
Dichmann DS, Walentek P, Harland RM. The Alternative Splicing Regulator Tra2b Is Required for Somitogenesis and Regulates Splicing of an Inhibitory Wnt11b Isoform. Cell Rep. 2015 Jan 21. pii: S2211-1247(14)01099-7. doi: 10.1016/j.celrep.2014.12.046. [Epub ahead of print].
http://www.ncbi.nlm.nih.gov/pubmed/25620705

Forcing a terminal intron inclusion:
Shchelkunova A, Ermolinsky B, Boyle M, Mendez I, Lehker M, Martirosyan KS, Kazansky AV. Tuning of Alternative Splicing - Switch From Proto-Oncogene to Tumor Suppressor. Int J Biol Sci. 2013;9(1):45-54. doi:10.7150/ijbs.5194.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3535533/

Blocking regulatory elements to increase intron retention:
Parra M, Yeo G, Conboy JG. Intron Retention Mechanisms That Regulate SF3B1 and Mitoferrin Gene Expression during Late Erythropoiesis. Blood. 2016;128:1200.
http://www.bloodjournal.org/content/128/22/1200

Presentations/posters from the 2020 MDA conference involving Morpholinos

Long-term Safety & Efficacy of Golodirsen in Male Patients with Duchenne Muscular Dystrophy (DMD) Amenable to Exon 53 Skipping
https://mdaconference.org/node/936

PMO-based miRNA site blocking oligo (SBO) mediated utrophin upregulation in mdx mice, a therapeutic approach for Duchenne Muscular Dystrophy (DMD)
https://mdaconference.org/node/1000

Safety of eteplirsen in Duchenne Muscular Dystrophy post-Cardiac transplantation
https://mdaconference.org/node/913

Open-Label Evaluation of Eteplirsen in Male Patients With Duchenne Muscular Dystrophy Amenable to Exon 51 Skipping vs Untreated Control: PROMOVI Trial
https://mdaconference.org/node/1088

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