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Gene Correction of Point Mutations Using PolyPurine Reverse Hoogsteen Hairpins Technology

Monogenic disorders are often the result of single point mutations in specific genes, leading to the production of non-functional proteins. Different blood disorders such as ß-thalassemia, sickle cell disease, hereditary spherocytosis, Fanconi anemia, and Hemophilia A and B are usually caused by poi...

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Autores principales: Félix, Alex J., Solé, Anna, Noé, Véronique, Ciudad, Carlos J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8525393/
https://www.ncbi.nlm.nih.gov/pubmed/34713221
http://dx.doi.org/10.3389/fgeed.2020.583577
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author Félix, Alex J.
Solé, Anna
Noé, Véronique
Ciudad, Carlos J.
author_facet Félix, Alex J.
Solé, Anna
Noé, Véronique
Ciudad, Carlos J.
author_sort Félix, Alex J.
collection PubMed
description Monogenic disorders are often the result of single point mutations in specific genes, leading to the production of non-functional proteins. Different blood disorders such as ß-thalassemia, sickle cell disease, hereditary spherocytosis, Fanconi anemia, and Hemophilia A and B are usually caused by point mutations. Gene editing tools including TALENs, ZFNs, or CRISPR/Cas platforms have been developed to correct mutations responsible for different diseases. However, alternative molecular tools such as triplex-forming oligonucleotides and their derivatives (e.g., peptide nucleic acids), not relying on nuclease activity, have also demonstrated their ability to correct mutations in the DNA. Here, we review the Repair-PolyPurine Reverse Hoogsteen hairpins (PPRHs) technology, which can represent an alternative gene editing tool within this field. Repair-PPRHs are non-modified single-stranded DNA molecules formed by two polypurine mirror repeat sequences linked by a five-thymidine bridge, followed by an extended sequence at one end of the molecule which is homologous to the DNA sequence to be repaired but containing the corrected nucleotide. The two polypurine arms of the PPRH are bound by intramolecular reverse-Hoogsteen bonds between the purines, thus forming a hairpin structure. This hairpin core binds to polypyrimidine tracts located relatively near the target mutation in the dsDNA in a sequence-specific manner by Watson-Crick bonds, thus producing a triplex structure which stimulates recombination. This technology has been successfully employed to repair a collection of mutants of the dhfr and aprt genes within their endogenous loci in mammalian cells and could be suitable for the correction of mutations responsible for blood disorders.
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spelling pubmed-85253932021-10-27 Gene Correction of Point Mutations Using PolyPurine Reverse Hoogsteen Hairpins Technology Félix, Alex J. Solé, Anna Noé, Véronique Ciudad, Carlos J. Front Genome Ed Genome Editing Monogenic disorders are often the result of single point mutations in specific genes, leading to the production of non-functional proteins. Different blood disorders such as ß-thalassemia, sickle cell disease, hereditary spherocytosis, Fanconi anemia, and Hemophilia A and B are usually caused by point mutations. Gene editing tools including TALENs, ZFNs, or CRISPR/Cas platforms have been developed to correct mutations responsible for different diseases. However, alternative molecular tools such as triplex-forming oligonucleotides and their derivatives (e.g., peptide nucleic acids), not relying on nuclease activity, have also demonstrated their ability to correct mutations in the DNA. Here, we review the Repair-PolyPurine Reverse Hoogsteen hairpins (PPRHs) technology, which can represent an alternative gene editing tool within this field. Repair-PPRHs are non-modified single-stranded DNA molecules formed by two polypurine mirror repeat sequences linked by a five-thymidine bridge, followed by an extended sequence at one end of the molecule which is homologous to the DNA sequence to be repaired but containing the corrected nucleotide. The two polypurine arms of the PPRH are bound by intramolecular reverse-Hoogsteen bonds between the purines, thus forming a hairpin structure. This hairpin core binds to polypyrimidine tracts located relatively near the target mutation in the dsDNA in a sequence-specific manner by Watson-Crick bonds, thus producing a triplex structure which stimulates recombination. This technology has been successfully employed to repair a collection of mutants of the dhfr and aprt genes within their endogenous loci in mammalian cells and could be suitable for the correction of mutations responsible for blood disorders. Frontiers Media S.A. 2020-10-29 /pmc/articles/PMC8525393/ /pubmed/34713221 http://dx.doi.org/10.3389/fgeed.2020.583577 Text en Copyright © 2020 Félix, Solé, Noé and Ciudad. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Genome Editing
Félix, Alex J.
Solé, Anna
Noé, Véronique
Ciudad, Carlos J.
Gene Correction of Point Mutations Using PolyPurine Reverse Hoogsteen Hairpins Technology
title Gene Correction of Point Mutations Using PolyPurine Reverse Hoogsteen Hairpins Technology
title_full Gene Correction of Point Mutations Using PolyPurine Reverse Hoogsteen Hairpins Technology
title_fullStr Gene Correction of Point Mutations Using PolyPurine Reverse Hoogsteen Hairpins Technology
title_full_unstemmed Gene Correction of Point Mutations Using PolyPurine Reverse Hoogsteen Hairpins Technology
title_short Gene Correction of Point Mutations Using PolyPurine Reverse Hoogsteen Hairpins Technology
title_sort gene correction of point mutations using polypurine reverse hoogsteen hairpins technology
topic Genome Editing
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8525393/
https://www.ncbi.nlm.nih.gov/pubmed/34713221
http://dx.doi.org/10.3389/fgeed.2020.583577
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