You are here

Jon-D's blog

A broader view: Approved Gene Therapy Products 2019

Development and Clinical Translation of Approved Gene Therapy Products for Genetic Disorders.

Shahryari1 A, Jazi MS, Mohammadi S, Nikoo HR, Nazari Z, Hosseini ES, Burtscher I, Mowla SJ, Lickert H.

Front Genet. 25 September 2019. doi:10.3389/fgene.2019.00868

https://www.frontiersin.org/articles/10.3389/fgene.2019.00868/full

Paper: Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase.

https://www.mdpi.com/1420-3049/24/18/3287

This paper describes using a microbial transglutamase to put an azide group in a peptide onto the terminal carboxylic acid of an antibody protein, then using Cu-free click chemistry to affix an antisense oligo at that site. They didn't use Morpholinos, but in principle one could.

Huggins IJ, Medina CA, Springer AD, van den Berg A, Jadhav S, Cui X, Dowdy SF. Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase. Molecules. 2019 Sep 10;24(18). pii: E3287. doi: 10.3390/molecules24183287.

Paper in delivery in cultures: In Vitro Validation of Phosphorodiamidate Morpholino Oligomers.

Aung-Htut MT, McIntosh CS, West KA, Fletcher S, Wilton SD. In Vitro Validation of Phosphorodiamidate Morpholino Oligomers. Molecules. 2019;24(6):2922 doi:0.3390/molecules24162922

https://www.mdpi.com/1420-3049/24/16/2922

cell culture: human dermal fibroblasts, human myoblasts
electroporation (nucelofection), Lipofectin, Lipofectamine, Endo-Porter

Delay in development and behavioural abnormalities in the absence of p53 in zebrafish

Delay in development and behavioural abnormalities in the absence of p53 in zebrafish.
Elabd S, Jabeen NA, Gerber V, Peravali R, Bourdon JC, Kancherla S, Vallone D, Blattner C.
PLoS One. 2019 Jul 19;14(7):e0220069. doi: 10.1371/journal.pone.0220069. eCollection 2019.
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0220069

This paper reports changes in developmental timing and behavior of p53-/- mutant embryos. While previous reports had suggested that p53 is dispensable for zebrafish development, the story might not be so simple. Previously a p53 knockdown was expected to produce an essentially wild-type embryo, but on the basis of this new work the results of Morpholino co-injection with p53 Morpholinos should be interpreted cautiously for contributions to the observed phenotype due to lack of p53. Still, given the gross abnormalities of an embryo in which the p53 apoptotic response is triggered by a knockdown, eliminating these abnormalities by a concurrent p53 knockdown is likely to remain a valuable technique for revealing gene function.

Example of specificity control using a Morpholino in its mutant

This paper is a nice example of Morpholino specificity control by using a Morpholino in its corresponding mutant.

Chambers BE, Gerlach GF, Clark EG, Chen KH, Levesque AE, Leshchiner I, Goessling W, Wingert RA. Tfap2a is a novel gatekeeper of nephron differentiation during kidney development. Development. 2019 Jun 3. pii: dev.172387. doi: 10.1242/dev.172387. [Epub ahead of print]

https://dev.biologists.org/content/146/13/dev172387

See especially figure S4:
http://dev.biologists.org/lookup/doi/10.1242/dev.172387.supplemental

Targeting mitochondrially-encoded circular RNAs in the cytosol: mitochondrial protein uptake suppression

Here is an interesting report of Morpholinos targeting mitochondrially-encoded circular RNAs in the cytosol to prevent RNA-chaperoned protein uptake into the mitochondria.

Liu X, Wang X, Li J, Hu S, Deng Y, Yin H, Bao X, Zhang QC, Wang G, Wang B, Shi Q, Shan G. The identification of mecciRNAs and their roles in mitochondrial entry of proteins. bioRxiv. 2019;[preprint] doi:10.1101/668665

Dosing mama mouse i.v. for transplacental delivery of Vivo-Morpholinos to pups

Touma M, Kang X, Gao F, Zhao Y, Cass AA, Biniwale R, Xiao X, Eghbali M, Coppola G, Reemtsen B, Wang Y. Wnt11 regulates cardiac chamber development and disease during perinatal maturation. JCI Insight. 2017 Sep 7;2(17). pii: 94904. doi: 10.1172/jci.insight.94904. eCollection 2017 Sep 7.

https://insight.jci.org/articles/view/94904

Review - Antisense-mediated splice intervention to treat human disease: the odyssey continues

Pitout I, Flynn LL, Wilton SD, Fletcher S.

Antisense-mediated splice intervention to treat human disease: the odyssey continues.

F1000Research 2019,8(F1000 Faculty Rev):710 doi:10.12688/f1000research.18466.1

https://f1000research.com/articles/8-710/v1

Innate immune stimulation by Morpholinos in Xenopus?

The discussion of innate immune stimulation by Morpholinos in Xenopus continues.

Paraiso KD, Blitz IL, Zhou JJ, Cho KWY. Morpholinos Do Not Elicit an Innate Immune Response during Early Xenopus Embryogenesis. Dev Cell. 2019;49(4):643-50. doi:10.1016/j.devcel.2019.04.019
https://www.cell.com/developmental-cell/fulltext/S1534-5807(19)30316-8

Gentsch GE, Spruce T, Owens NDL, Monteiro RS, Smith JC. The Innate Immune Response of Frog Embryos to Antisense Morpholino Oligomers Depends on Developmental Stage, GC Content and Dose. Dev Cell. 2019;49(4):506-7. doi:10.1016/j.devcel.2019.05.004
https://www.cell.com/developmental-cell/fulltext/S1534-5807(19)30375-2

These new papers extend the work presented in:
Gentsch GE, Spruce T, Monteiro RS, Owens NDL, Martin SR, Smith JC. Innate Immune Response and Off-Target Mis-splicing Are Common Morpholino-Induced Side Effects in Xenopus. Dev Cell. 2018;[Epub ahead of print] doi:10.1016/j.devcel.2018.01.022.
http://www.cell.com/developmental-cell/fulltext/S1534-5807(18)30057-1

Pages

Subscribe to RSS - Jon-D's blog