scholarly journals Novel Disulfide-Bridged Bioresponsive Antisense Oligonucleotide Induces Efficient Splice Modulation in Muscle Myotubes in Vitro

ACS Omega ◽  
2020 ◽  
Vol 5 (29) ◽  
pp. 18035-18039
Author(s):  
Bao T. Le ◽  
Tamer R. Kosbar ◽  
Rakesh N. Veedu
Keyword(s):  
Antisense Oligonucleotide

scholarly journals Ethylcellulose nanoparticles as a new “in vitro” transfection tool for antisense oligonucleotide delivery

2020 ◽  
Vol 229 ◽  
pp. 115451 ◽  
Author(s):  
S. Leitner ◽  
S. Grijalvo ◽  
C. Solans ◽  
R. Eritja ◽  
M.J. García-Celma ◽  
...  
Keyword(s):  
Antisense Oligonucleotide ◽  
Oligonucleotide Delivery ◽  
In Vitro Transfection

scholarly journals Muscle-specific microRNA miR-206 promotes muscle differentiation

2006 ◽  
Vol 174 (5) ◽  
pp. 677-687 ◽  
Author(s):  
Hak Kyun Kim ◽  
Yong Sun Lee ◽  
Umasundari Sivaprasad ◽  
Ankit Malhotra ◽  
Anindya Dutta
Keyword(s):  
Antisense Oligonucleotide ◽  
Small Interfering Rna ◽  
Degradation Process ◽  
Microrna Target ◽  
C2c12 Myoblasts ◽  
Target Sites ◽  
The Many ◽  
Interfering Rna ◽  
In Vitro Transfection

Three muscle-specific microRNAs, miR-206, -1, and -133, are induced during differentiation of C2C12 myoblasts in vitro. Transfection of miR-206 promotes differentiation despite the presence of serum, whereas inhibition of the microRNA by antisense oligonucleotide inhibits cell cycle withdrawal and differentiation, which are normally induced by serum deprivation. Among the many mRNAs that are down-regulated by miR-206, the p180 subunit of DNA polymerase α and three other genes are shown to be direct targets. Down-regulation of the polymerase inhibits DNA synthesis, an important component of the differentiation program. The direct targets are decreased by mRNA cleavage that is dependent on predicted microRNA target sites. Unlike small interfering RNA–directed cleavage, however, the 5′ ends of the cleavage fragments are distributed and not confined to the target sites, suggesting involvement of exonucleases in the degradation process. In addition, inhibitors of myogenic transcription factors, Id1-3 and MyoR, are decreased upon miR-206 introduction, suggesting the presence of additional mechanisms by which microRNAs enforce the differentiation program.

The uptake of a fluorescently labelled antisense oligonucleotide in vitro and in vivo

2005 ◽  
Vol 147 (1) ◽  
pp. 48-54 ◽  
Author(s):  
Emma S.J. Robinson ◽  
David J. Nutt ◽  
Helen C. Jackson ◽  
Alan L. Hudson
Keyword(s):  
Antisense Oligonucleotide

scholarly journals Magnetofection—A highly efficient tool for antisense oligonucleotide delivery in vitro and in vivo

2003 ◽  
Vol 7 (5) ◽  
pp. 700-710 ◽  
Author(s):  
Florian Krötz ◽  
Cor de Wit ◽  
Hae-Young Sohn ◽  
Stefan Zahler ◽  
Torsten Gloe ◽  
...  
Keyword(s):  
Antisense Oligonucleotide ◽  
Efficient Tool ◽  
Highly Efficient ◽  
Oligonucleotide Delivery

An anionic long-circulating liposome that improves radioiodinated antisense oligonucleotide delivery in vitro and in vivo

2011 ◽  
Vol 31 (1) ◽  
pp. 20-28 ◽  
Author(s):  
Jiawei Xie ◽  
Chao Ma ◽  
Jizheng Lin ◽  
Guoming Wang ◽  
Anren Kuang ◽  
...  
Keyword(s):  
Antisense Oligonucleotide ◽  
Oligonucleotide Delivery

scholarly journals Dual Fluorescence Splicing Reporter Minigene Identifies an Antisense Oligonucleotide to Skip Exon v8 of the CD44 Gene

2020 ◽  
Vol 21 (23) ◽  
pp. 9136
Author(s):  
Sachiyo Fukushima ◽  
Manal Farea ◽  
Kazuhiro Maeta ◽  
Abdul Qawee Mahyoob Rani ◽  
Kazumichi Fujioka ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Antisense Oligonucleotide ◽  
High Throughput Screening ◽  
Pcr Amplification ◽  
Dual Fluorescence ◽  
Reverse Transcription Pcr ◽  
In Vitro Splicing ◽  
Split Egfp ◽  
Artificial Intron

Splicing reporter minigenes are used in cell-based in vitro splicing studies. Exon skippable antisense oligonucleotide (ASO) has been identified using minigene splicing assays, but these assays include a time- and cost-consuming step of reverse transcription PCR amplification. To make in vitro splicing assay easier, a ready-made minigene (FMv2) amenable to quantitative splicing analysis by fluorescence microscopy was constructed. FMv2 was designed to encode two fluorescence proteins namely, mCherry, a transfection marker and split eGFP, a marker of splicing reaction. The split eGFP was intervened by an artificial intron containing a multicloning site sequence. Expectedly, FMv2 transfected HeLa cells produced not only red mCherry but also green eGFP signals. Transfection of FMv2CD44v8, a modified clone of FMv2 carrying an insertion of CD44 exon v8 in the multicloning site, that was applied to screen exon v8 skippable ASO, produced only red signals. Among seven different ASOs tested against exon v8, ASO#14 produced the highest index of green signal positive cells. Hence, ASO#14 was the most efficient exon v8 skippable ASO. Notably, the well containing ASO#14 was clearly identified among the 96 wells containing randomly added ASOs, enabling high throughput screening. A ready-made FMv2 is expected to contribute to identify exon skippable ASOs.

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