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Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing

Clustered regularly interspaced short palindromic repeat (CRISPR) and their associated protein (Cas-9) is the most effective, efficient, and accurate method of genome editing tool in all living cells and utilized in many applied disciplines. Guide RNA (gRNA) and CRISPR-associated (Cas-9) proteins ar...

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Autores principales: Asmamaw, Misganaw, Zawdie, Belay
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8388126/
https://www.ncbi.nlm.nih.gov/pubmed/34456559
http://dx.doi.org/10.2147/BTT.S326422
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author Asmamaw, Misganaw
Zawdie, Belay
author_facet Asmamaw, Misganaw
Zawdie, Belay
author_sort Asmamaw, Misganaw
collection PubMed
description Clustered regularly interspaced short palindromic repeat (CRISPR) and their associated protein (Cas-9) is the most effective, efficient, and accurate method of genome editing tool in all living cells and utilized in many applied disciplines. Guide RNA (gRNA) and CRISPR-associated (Cas-9) proteins are the two essential components in CRISPR/Cas-9 system. The mechanism of CRISPR/Cas-9 genome editing contains three steps, recognition, cleavage, and repair. The designed sgRNA recognizes the target sequence in the gene of interest through a complementary base pair. While the Cas-9 nuclease makes double-stranded breaks at a site 3 base pair upstream to protospacer adjacent motif, then the double-stranded break is repaired by either non-homologous end joining or homology-directed repair cellular mechanisms. The CRISPR/Cas-9 genome-editing tool has a wide number of applications in many areas including medicine, agriculture, and biotechnology. In agriculture, it could help in the design of new grains to improve their nutritional value. In medicine, it is being investigated for cancers, HIV, and gene therapy such as sickle cell disease, cystic fibrosis, and Duchenne muscular dystrophy. The technology is also being utilized in the regulation of specific genes through the advanced modification of Cas-9 protein. However, immunogenicity, effective delivery systems, off-target effect, and ethical issues have been the major barriers to extend the technology in clinical applications. Although CRISPR/Cas-9 becomes a new era in molecular biology and has countless roles ranging from basic molecular researches to clinical applications, there are still challenges to rub in the practical applications and various improvements are needed to overcome obstacles.
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spelling pubmed-83881262021-08-27 Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing Asmamaw, Misganaw Zawdie, Belay Biologics Review Clustered regularly interspaced short palindromic repeat (CRISPR) and their associated protein (Cas-9) is the most effective, efficient, and accurate method of genome editing tool in all living cells and utilized in many applied disciplines. Guide RNA (gRNA) and CRISPR-associated (Cas-9) proteins are the two essential components in CRISPR/Cas-9 system. The mechanism of CRISPR/Cas-9 genome editing contains three steps, recognition, cleavage, and repair. The designed sgRNA recognizes the target sequence in the gene of interest through a complementary base pair. While the Cas-9 nuclease makes double-stranded breaks at a site 3 base pair upstream to protospacer adjacent motif, then the double-stranded break is repaired by either non-homologous end joining or homology-directed repair cellular mechanisms. The CRISPR/Cas-9 genome-editing tool has a wide number of applications in many areas including medicine, agriculture, and biotechnology. In agriculture, it could help in the design of new grains to improve their nutritional value. In medicine, it is being investigated for cancers, HIV, and gene therapy such as sickle cell disease, cystic fibrosis, and Duchenne muscular dystrophy. The technology is also being utilized in the regulation of specific genes through the advanced modification of Cas-9 protein. However, immunogenicity, effective delivery systems, off-target effect, and ethical issues have been the major barriers to extend the technology in clinical applications. Although CRISPR/Cas-9 becomes a new era in molecular biology and has countless roles ranging from basic molecular researches to clinical applications, there are still challenges to rub in the practical applications and various improvements are needed to overcome obstacles. Dove 2021-08-21 /pmc/articles/PMC8388126/ /pubmed/34456559 http://dx.doi.org/10.2147/BTT.S326422 Text en © 2021 Asmamaw and Zawdie. https://creativecommons.org/licenses/by-nc/3.0/This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/ (https://creativecommons.org/licenses/by-nc/3.0/) ). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php).
spellingShingle Review
Asmamaw, Misganaw
Zawdie, Belay
Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing
title Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing
title_full Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing
title_fullStr Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing
title_full_unstemmed Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing
title_short Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing
title_sort mechanism and applications of crispr/cas-9-mediated genome editing
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8388126/
https://www.ncbi.nlm.nih.gov/pubmed/34456559
http://dx.doi.org/10.2147/BTT.S326422
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