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|Time for some music! Evo-devo||Wednesday, September 27, 2017 - 11:47|
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|
|Review of eteplirsen for DMD||
Mutation-Based Therapy for Duchenne Muscular Dystrophy: Antisense Treatment Arrives in the Clinic.
|Monday, September 11, 2017 - 14:04|
|BLAST homology and specificity controls||
I have had many discussions about specificity with a Morpholino user who does very careful BLAST analysis of each oligo prior to ordering. I had previously suggested that about 14 bases of complementary (without significant flanking complementarity) is probably an acceptable level of stability for off-target interactions found by BLAST. However, in light of a new report I agreed that a 13-base complementarity at a splice junction is enough to raise concern. Here is an excerpt of my response to her.
|Wednesday, August 2, 2017 - 08:21|
|Paper on Morpholino specificity and CRISPR-MO combination||
Number of inadvertent RNA targets for morpholino knockdown in Danio rerio is largely underestimated: evidence from the study of Ser/Arg-rich splicing factors.
Joris M, Schloesser M, Baurain D, Hanikenne M, Muller M, Motte P.
Nucleic Acids Res. 2017;[Epub ahead of print] doi:10.1093/nar/gkx638.
|Tuesday, August 1, 2017 - 14:35|
|Formatting sequence information for emails (splice modifying oligo designs)||
To email sequence information for design of a splice-modifying Morpholino, write introns in lower case and exons in upper case and write the sense sequence 5'-3'. If you show a start codon, put parentheses around it. Here is an example:
All I need for the design are 10 exonic and 25 intronic bases, like this:
|Tuesday, July 25, 2017 - 08:02|
|Genetic compensation: A phenomenon in search of mechanisms||
Genetic compensation: A phenomenon in search of mechanisms.
El-Brolosy MA, Stainier DYR.
PLoS Genet. 2017 Jul 13;13(7):e1006780. doi: 10.1371/journal.pgen.1006780. eCollection 2017 Jul. Review.
|Friday, July 14, 2017 - 10:29|
|Book: Morpholino Oligomers||
Hong and I edited a book: Morpholino Oligomers. 2017. Methods in Molecular Biology. Volume 1565. Humana Press (Springer). doi:10.1007/978-1-4939-6817-6
Links to chapters follow.
1. Summerton JE. Invention and Early History of Morpholinos: From Pipe Dream to Practical Products. Methods Mol Biol. 2017;1565:1-15. doi: 10.1007/978-1-4939-6817-6_1. Abstract: PubMed
|Tuesday, April 11, 2017 - 01:57|
|Review of FDA-approved antisense (2017)||
FDA-Approved Oligonucleotide Therapies in 2017.
|Thursday, April 6, 2017 - 08:40|
|Using Morpholinos to Control Gene Expression (in CPNAC)||
Wiley has published an update to my Morpholino how-to and background review.
Moulton JD. Using Morpholinos to Control Gene Expression. Curr Protoc Nucleic Acid Chem. 2017 Mar 2;68:4.30.1-4.30.29. doi: 10.1002/cpnc.21.
|Saturday, March 4, 2017 - 03:26|