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APC Is an RNA-Binding Protein, and Its Interactome Provides a Link to Neural Development and Microtubule Assembly

Preitner N, Quan J, Nowakowski DW, Hancock ML, Shi J, Tcherkezian J, Young-Pearse TL, Flanagan JG. APC Is an RNA-Binding Protein, and Its Interactome Provides a Link to Neural Development and Microtubule Assembly. Cell. 2014 Jul 17;158(2):368-82. doi: 10.1016/j.cell.2014.05.042.

This paper reports rat brain in utero injection then electroporation of Morpohlinos as well as mouse experiments with peptide nucleic acids.

Tuesday, July 22, 2014 - 09:50
Oligos targeted to trigger nonsense-mediated decay (NMD)

This paper provides some experimental support for targeting steric-blocking oligos to alter splicing and trigger nonsense-mediated decay. The paper compared uniform 2'-methoxyethyl phosphorothioate oligos with 2'-MOE phosphorothioate gapmers (steric blocking versus RNase-H competent, respectively).

Nonsense-mediated decay as a terminating mechanism for antisense oligonucleotides.
Ward AJ, Norrbom M, Chun S, Bennett CF, Rigo F.
Nucleic Acids Res. 2014 May 1;42(9):5871-9. doi: 10.1093/nar/gku184. Epub 2014 Mar 3.

Thursday, June 12, 2014 - 15:38
Intranasal administration in mice of peptide-conjugated Morpholinos

Peptide-conjugated Morpholinos were administered intranasally to mice to alter the immune response to subsequent intranasal influenza doses. "BALB/c mice were anesthetized with ketamine, followed by i.n. administration with 100 mg (approximately 5 mg/kg) PPMO (in 40 ml of PBS)." Repeat dosing at 24 hours, influenza inoculation at 48 hours.

Monday, June 2, 2014 - 12:09
Knockdowns in mosquito guts

Vivo-Morpholinos were fed to mosquitos. The oligos decreased the amount of MAPK protein in the midgut and the number of Plasmodium oocysts in the mudgut.

"The feeding protocol described in the present study provides several advantages over current methods for gene knockdown in insects. The method is easy to apply, requires no special skills for delivery, and is highly target specific. ... This protocol should therefore be considered as an efficient alternative in studies requiring target specific protein knockdown in mosquitoes and other arthropods."

Thursday, May 29, 2014 - 11:45
Morpholino electroporated into human T-cells induces alternative splicing without interferon response Michel M, Wilhelmi I, Schultz A-S, Preussner M, Heyd F. Activation-induced Tumor Necrosis Factor Receptor-associated Factor 3 (Traf3) Alternative Splicing Controls the Noncanonical Nuclear Factor κB Pathway and Chemokine Expression in Human T Cells. J Biol Chem. 2014;289:13651-60. doi:10.1074/jbc.M113.526269

"Furthermore, the interferon response, which can be regulated by Traf3 (35), was not altered in Traf3E8 MO-treated cells (data not shown)" supports that the Morpholino did not induce an innate immune response.

Friday, May 9, 2014 - 09:21
Vivo-Morpholino in rat brain Shen H-w, Scofield MD, Boger H, Hensley M, Kalivas PW. Synaptic Glutamate Spillover Due to Impaired Glutamate Uptake Mediates Heroin Relapse. J Neurosci. 2014;34(16):5649-57 doi:10.1523/JNEUROSCI.4564-13.2014

http://www.jneurosci.org/content/34/16/5649.abstract

Thursday, April 17, 2014 - 09:16
Nanoparticle activated by near-IR delivers and cleaves Photo-Morpholino Jayakumar MK, Bansal A, Huang K, Yao R, Li BN, Zhang Y. Near-Infrared-Light-Based Nano-Platform Boosts Endosomal Escape and Controls Gene Knockdown in Vivo. ACS Nano. 2014 Apr 14. [Epub ahead of print]

http://www.cell.com/current-biology/abstract/S0960-9822%2812%2901327-9

Wednesday, April 16, 2014 - 13:34
Using a GFP reporter to test a knockdown Romaker D, Kumar V, Cerqueira DM, Cox RM, Wessely O. MicroRNAs are critical regulators of tuberous sclerosis complex and mTORC1 activity in the size control of the Xenopus kidney. Proc Natl Acad Sci U S A. 2014 Apr 14. [Epub ahead of print]

The authors report several test constructs in which their upstream Morpholino target is ligated to a GFP coding sequence and then tested for its response to a Morpholino translation blocker. See figure S6 in the supporting online information.

Wednesday, April 16, 2014 - 12:59
Jon's blog for notes on new applications and pointers to techniques Welcome! I'll be keeping notes here on citations that demonstrate techniques that catch my interest using Morpholino oligos. Wednesday, April 16, 2014 - 12:00

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