scholarly journals miR-7 Knockdown by Peptide Nucleic Acids in the Ascidian Ciona intestinalis

2019 ◽  
Vol 20 (20) ◽  
pp. 5127 ◽  
Author(s):  
Silvia Mercurio ◽  
Silvia Cauteruccio ◽  
Raoul Manenti ◽  
Simona Candiani ◽  
Giorgio Scarì ◽  
...  

Peptide Nucleic Acids (PNAs) are synthetic mimics of natural oligonucleotides, which bind complementary DNA/RNA strands with high sequence specificity. They display numerous advantages, but in vivo applications are still rare. One of the main drawbacks of PNAs application is the poor cellular uptake that could be overcome by using experimental models, in which microinjection techniques allow direct delivery of molecules into eggs. Thus, in this communication, we investigated PNAs efficiency in miR-7 downregulation and compared its effects with those obtained with the commercially available antisense molecule, Antagomir (Dharmacon) in the ascidian Ciona intestinalis. Ascidians are marine invertebrates closely related to vertebrates, in which PNA techniques have not been applied yet. Our results suggested that anti-miR-7 PNAs were able to reach their specific targets in the developing ascidian embryos with high efficiency, as the same effects were obtained with both PNA and Antagomir. To the best of our knowledge, this is the first evidence that unmodified PNAs can be applied in in vivo knockdown strategies when directly injected into eggs.

2020 ◽  
Vol 21 (6) ◽  
pp. 2001
Author(s):  
Silvia Mercurio ◽  
Silvia Cauteruccio ◽  
Raoul Manenti ◽  
Simona Candiani ◽  
Giorgio Scarì ◽  
...  

The microRNAs are small RNAs that regulate gene expression at the post-transcriptional level and can be involved in the onset of neurodegenerative diseases and cancer. They are emerging as possible targets for antisense-based therapy, even though the in vivo stability of miRNA analogues is still questioned. We tested the ability of peptide nucleic acids, a novel class of nucleic acid mimics, to downregulate miR-9 in vivo in an invertebrate model organism, the ascidian Ciona intestinalis, by microinjection of antisense molecules in the eggs. It is known that miR-9 is a well-conserved microRNA in bilaterians and we found that it is expressed in epidermal sensory neurons of the tail in the larva of C. intestinalis. Larvae developed from injected eggs showed a reduced differentiation of tail neurons, confirming the possibility to use peptide nucleic acid PNA to downregulate miRNA in a whole organism. By identifying putative targets of miR-9, we discuss the role of this miRNA in the development of the peripheral nervous system of ascidians.


Life Sciences ◽  
2002 ◽  
Vol 71 (3) ◽  
pp. 325-337 ◽  
Author(s):  
Beth M. McMahon ◽  
Jennifer A. Stewart ◽  
M.D. Bitner ◽  
Abdul Fauq ◽  
Daniel J. McCormick ◽  
...  

2006 ◽  
Vol 71 (7) ◽  
pp. 929-955 ◽  
Author(s):  
Vladimir A. Efimov ◽  
Oksana G. Chakhmakhcheva

With the aim to improve physicochemical and biological properties of natural oligonucleotides, many types of DNA analogues and mimics are designed on the basis of hydroxyproline and its derivatives, and their properties are evaluated. Among them, two types of DNA mimics representing hetero-oligomers constructed from alternating monomers of phosphono peptide nucleic acids and monomers on the base of trans-1-acetyl-4-hydroxy-L-proline (HypNA-pPNAs) and oligomers constructed from monomers containing (2S,4R)-1-acetyl-4-hydroxypyrrolidine-2-phosphonic acid backbone (pHypNAs) are of particular interest. In a set of in vitro and in vivo assays, it was shown that HypNA-pPNAs and pHypNAs demonstrated a high potential for the use in nucleic acid based diagnostics, isolation of nucleic acids and antisense experiments. A review with 53 references.


FEBS Letters ◽  
1998 ◽  
Vol 421 (3) ◽  
pp. 280-284 ◽  
Author(s):  
Beth M Tyler ◽  
Daniel J McCormick ◽  
Clark V Hoshall ◽  
Christopher L Douglas ◽  
Karen Jansen ◽  
...  

2019 ◽  
Vol 16 (10) ◽  
pp. 4223-4229 ◽  
Author(s):  
Chenxia M. Guan ◽  
Alyssa B. Chinen ◽  
Jennifer R. Ferrer ◽  
Caroline H. Ko ◽  
Chad A. Mirkin

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Nhan Huynh ◽  
Noah Depner ◽  
Raegan Larson ◽  
Kirst King-Jones

AbstractAdvances in CRISPR technology have immensely improved our ability to manipulate nucleic acids, and the recent discovery of the RNA-targeting endonuclease Cas13 adds even further functionality. Here, we show that Cas13 works efficiently in Drosophila, both ex vivo and in vivo. We test 44 different Cas13 variants to identify enzymes with the best overall performance and show that Cas13 could target endogenous Drosophila transcripts in vivo with high efficiency and specificity. We also develop Cas13 applications to edit mRNAs and target mitochondrial transcripts. Our vector collection represents a versatile tool collection to manipulate gene expression at the post-transcriptional level.


2010 ◽  
Vol 38 (13) ◽  
pp. 4466-4475 ◽  
Author(s):  
Martin M. Fabani ◽  
Cei Abreu-Goodger ◽  
Donna Williams ◽  
Paul A. Lyons ◽  
Adrian G. Torres ◽  
...  

2004 ◽  
Vol 12 (6) ◽  
pp. 363-372 ◽  
Author(s):  
E. Wickstrom ◽  
M. Choob ◽  
K.A. Urtishak ◽  
X. Tian ◽  
N. Sternheim ◽  
...  

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