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Jon's Blog
| Post date | ||
|---|---|---|
| Skipping Multiple Exons: Review of DMD Morpholino, Vivo-Morpholino & PPMO work |
Review of DMD multiple exon skipping with Morpholino, Vivo-Morpholino & PPMO by Toshifumi Yokota's group. Skipping Multiple Exons to Treat DMD-Promises and Challenges. Aslesh T, Maruyama R, Yokota T. Biomedicines. 2018 Jan 2;6(1). pii: E1. doi: 10.3390/biomedicines6010001. Review. |
Monday, January 8, 2018 - 07:46 |
| Morpholino-based skipping of an exon caused inclusion of another exon, following that transcript's pattern of normal alternative splicing. |
Here is an example of an unusual outcome from modifying splicing. Vivo-Morpholinos targeting exon 9 caused skipping of exon 9 and inclusion of exon 10. In normal alternative splicing of this RNA, either exon 9 is included and exon 10 is excised (Pkm1) or exon 9 is excised and exon 10 is excluded (Pkm2). |
Wednesday, December 20, 2017 - 12:00 |
| Gel retardation assay using RNA with a complementary Morpholino |
This image is of a gel retardation assay using RNA with a complementary Morpholino altering its migration. https://static-content.springer.com/esm/art%3A10.1007%2Fs00705-017-3666-... |
Tuesday, December 19, 2017 - 14:15 |
| Paper: Hybrid splicing minigene and antisense oligonucleotides as efficient tools to determine functional protein/RNA interactions |
This paper describes setting up systems to determine splicing-related protein-RNA interactions; this might be especially useful for confirming whether a particular putatuve splice-regulatory protein binding site is actually involved in modulating splicing. The oligos reported were not Morpholinos, but I expect this system would be compatible with Morpohlinos. Hybrid splicing minigene and antisense oligonucleotides as efficient tools to determine functional protein/RNA interactions. Cywoniuk P, Taylor K, Sznajder ŁJ, Sobczak K. |
Monday, December 18, 2017 - 09:18 |
| Chemistry, mechanism and clinical status of antisense oligonucleotides and duplex RNAs |
This is an overview of various antisense drugs, approved, withdrawn, and in clinical trials, along with an introduction to various intisense structural types and modes of action. Shen X, Corey DR. Chemistry, mechanism and clinical status of antisense oligonucleotides and duplex RNAs. Nucleic Acid Research. 2017;[Epub ahead of print] doi:/10.1093/nar/gkx1239 |
Tuesday, December 12, 2017 - 08:42 |
| Impurities in Oligonucleotide Drug Substances and Drug Products |
Here's an interesting article on oligo drug impurities, written from a phosphorothioate perspective but with many ideas potentially applicable to Morpholino GMP production. |
Tuesday, December 12, 2017 - 08:36 |
| Why use 0.1N HCl to measure Morpholino concentration by UV spec? |
Here is an excerpt from a letter Jim Summerton recently wrote in which he described the reason why Morpholinos are quantitated by measuring UV absorbance at 265 nm in 0.1N HCl. ------------------------------------------------------------------------------------ |
Tuesday, November 28, 2017 - 09:45 |
| Targeting internal exon caused intron inclusion: example of a less-common outcome |
Here is an example where targeting an internal exon of a pre-mRNA caused inclusion of the adjacent intron, an unusual outcome. Vierstraete J, Willaert A, Vermassen P, Coucke PJ, Vral A, Claes KBM. Accurate quantification of homologous recombination in zebrafish: brca2 deficiency as a paradigm. Sci Rep. 2017;7:16518. doi:10.1038/s41598-017-16725-3 |
Tuesday, November 28, 2017 - 08:27 |
| Combining Vivo-Morpholinos & Endo-Porter in a hard-to-transfect cell line |
These investigators report enhanced delivery of Vivo-Morpholinos by combining them with Endo-Porter to deliver the oligos into a hard-to-transfect cell line. Adamo P, Porazinski S, Rajatileka S, Jumbe S, Hagen R, Cheung MK, Wilson I, Ladomery MR. The oncogenic transcription factor ERG represses the transcription of the tumour suppressor gene PTEN in prostate cancer cells. Oncol Lett. 2017 Nov;14(5):5605-5610. doi: 10.3892/ol.2017.6841. Epub 2017 Aug 28. |
Monday, November 27, 2017 - 11:23 |
| On the difficulty of making a good null mutant |
http://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.10... Anderson JL, Mulligan TS, Shen M-C, Wang H, Scahill CM, Tan FJ, Du SJ, Busch-Nentwich EM, Farber SA. mRNA processing in mutant zebrafish lines generated by chemical and CRISPR-mediated mutagenesis produces unexpected transcripts that escape nonsense-mediated decay. PLoS Genet. 2017;13(11):e1007105. doi:10.1371/journal.pgen.1007105 |
Tuesday, November 21, 2017 - 12:44 |
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