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Clickable PNA Probes for Imaging Human Telomeres and Poly(A) RNAs

[Image: see text] The ability to bind strongly to complementary nucleic acid sequences, invade complex nucleic acid structures, and resist degradation by cellular enzymes has made peptide nucleic acid (PNA) oligomers as very useful hybridization probes in molecular diagnosis. For such applications,...

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Autores principales: Sabale, Pramod M., Ambi, Uddhav B., Srivatsan, Seergazhi G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2018
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6289544/
https://www.ncbi.nlm.nih.gov/pubmed/30556003
http://dx.doi.org/10.1021/acsomega.8b02550
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author Sabale, Pramod M.
Ambi, Uddhav B.
Srivatsan, Seergazhi G.
author_facet Sabale, Pramod M.
Ambi, Uddhav B.
Srivatsan, Seergazhi G.
author_sort Sabale, Pramod M.
collection PubMed
description [Image: see text] The ability to bind strongly to complementary nucleic acid sequences, invade complex nucleic acid structures, and resist degradation by cellular enzymes has made peptide nucleic acid (PNA) oligomers as very useful hybridization probes in molecular diagnosis. For such applications, the PNA oligomers have to be labeled with appropriate reporters as they lack intrinsic labels that can be used in biophysical assays. Although solid-phase synthesis is commonly used to attach reporters onto PNA, development of milder and modular labeling methods will provide access to PNA oligomers labeled with a wider range of biophysical tags. Here, we describe the establishment of a postsynthetic modification strategy based on bioorthogonal chemical reactions in functionalizing PNA oligomers in solution with a variety of tags. A toolbox composed of alkyne- and azide-modified monomers were site-specifically incorporated into PNA oligomers and postsynthetically click-functionalized with various tags, ranging from sugar, amino acid, biotin, to fluorophores, by using copper(I)-catalyzed azide–alkyne cycloaddition, strain-promoted azide–alkyne cycloaddition, and Staudinger ligation reactions. As a proof of utility of this method, fluorescent PNA hybridization probes were developed and used in imaging human telomeres in chromosomes and poly(A) RNAs in cells. Taken together, this simple approach of generating a wide range of functional PNA oligomers will expand the use of PNA in molecular diagnosis.
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spelling pubmed-62895442018-12-12 Clickable PNA Probes for Imaging Human Telomeres and Poly(A) RNAs Sabale, Pramod M. Ambi, Uddhav B. Srivatsan, Seergazhi G. ACS Omega [Image: see text] The ability to bind strongly to complementary nucleic acid sequences, invade complex nucleic acid structures, and resist degradation by cellular enzymes has made peptide nucleic acid (PNA) oligomers as very useful hybridization probes in molecular diagnosis. For such applications, the PNA oligomers have to be labeled with appropriate reporters as they lack intrinsic labels that can be used in biophysical assays. Although solid-phase synthesis is commonly used to attach reporters onto PNA, development of milder and modular labeling methods will provide access to PNA oligomers labeled with a wider range of biophysical tags. Here, we describe the establishment of a postsynthetic modification strategy based on bioorthogonal chemical reactions in functionalizing PNA oligomers in solution with a variety of tags. A toolbox composed of alkyne- and azide-modified monomers were site-specifically incorporated into PNA oligomers and postsynthetically click-functionalized with various tags, ranging from sugar, amino acid, biotin, to fluorophores, by using copper(I)-catalyzed azide–alkyne cycloaddition, strain-promoted azide–alkyne cycloaddition, and Staudinger ligation reactions. As a proof of utility of this method, fluorescent PNA hybridization probes were developed and used in imaging human telomeres in chromosomes and poly(A) RNAs in cells. Taken together, this simple approach of generating a wide range of functional PNA oligomers will expand the use of PNA in molecular diagnosis. American Chemical Society 2018-11-12 /pmc/articles/PMC6289544/ /pubmed/30556003 http://dx.doi.org/10.1021/acsomega.8b02550 Text en Copyright © 2018 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes.
spellingShingle Sabale, Pramod M.
Ambi, Uddhav B.
Srivatsan, Seergazhi G.
Clickable PNA Probes for Imaging Human Telomeres and Poly(A) RNAs
title Clickable PNA Probes for Imaging Human Telomeres and Poly(A) RNAs
title_full Clickable PNA Probes for Imaging Human Telomeres and Poly(A) RNAs
title_fullStr Clickable PNA Probes for Imaging Human Telomeres and Poly(A) RNAs
title_full_unstemmed Clickable PNA Probes for Imaging Human Telomeres and Poly(A) RNAs
title_short Clickable PNA Probes for Imaging Human Telomeres and Poly(A) RNAs
title_sort clickable pna probes for imaging human telomeres and poly(a) rnas
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6289544/
https://www.ncbi.nlm.nih.gov/pubmed/30556003
http://dx.doi.org/10.1021/acsomega.8b02550
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