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Jon's Blog
| Post date | ||
|---|---|---|
| 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 |
| ON-based drugs approved by the FDA |
Here they are so far, the approved oligonucleotide drugs. Note, however, that Morpholinos are oligos but not nucleotides; Exondys 51 (eteplirsen) is the Morpholino drug. Table 1. ON-based drugs approved by the FDA. |
Tuesday, October 24, 2017 - 11:31 |
| Guidelines for morpholino use in zebrafish |
If you work with zebrafish or Morpholinos, please read this. Stainier DYR, Raz E, Lawson ND, Ekker SC, Burdine RD, Eisen JS, Ingham PW, Schulte-Merker S, Yelon D, Weinstein BM, Mullins MC, Wilson SW, Ramakrishnan L, Amacher SL, Neuhauss SCF, Meng A, Mochizuki N, Panula P, Moens CB. Guidelines for morpholino use in zebrafish. PLoS Genet. 2017 Oct 19;13(10):e1007000. doi: 10.1371/journal.pgen.1007000. eCollection 2017 Oct. No abstract available. |
Friday, October 20, 2017 - 08:31 |
| Mechanism of uptake of Morpholino oligos in dystrophic (DMD) muscle |
This work elucidates the mechanism of uptake of Morpholino oligos in dystrophic (DMD) muscle, including roles for muscle satellite cells and macrophages. Novak JS, Hogarth MW, Boehler JF, Nearing M, Vila MC, Heredia R, Fiorillo AA, Zhang A, Hathout Y, Hoffman EP, Jaiswal JK, Nagaraju K, Cirak S, Partridge TA. Nat Commun. 2017 Oct 16;8(1):941. doi: 10.1038/s41467-017-00924-7. |
Thursday, October 19, 2017 - 10:36 |
| Time for some music! Evo-devo |
Evo-Devo |
Wednesday, September 27, 2017 - 11:47 |
| Confirming Specificity |
There are several commonly-used strategies for confirming specificity of a Morpholino oligo. These are the five-mispair experiment, the second non-overlapping 5'-UTR oligo experiment (and the similar two oligos against one mRNA experiments), the mRNA rescue experiment, and my current favorite, the Morpholino-in-a-null-mutant experiment. |
Wednesday, September 27, 2017 - 08:46 |
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