CRISPR/Cas9 gene editing: a novel strategy for fighting drug resistance in respiratory disorders

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Abstract

Respiratory disorders are among the conditions that affect the respiratory system. The healthcare sector faces challenges due to the emergence of drug resistance to prescribed medications for these illnesses. However, there is a technology called CRISPR/Cas9, which uses RNA to guide DNA targeting. This technology has revolutionized our ability to manipulate and visualize the genome, leading to advancements in research and treatment development. It can effectively reverse epigenetic alterations that contribute to drug resistance. Some studies focused on health have shown that targeting genes using CRISPR/Cas9 can be challenging when it comes to reducing drug resistance in patients with respiratory disorders. Nevertheless, it is important to acknowledge the limitations of this technology, such as off-target effects, immune system reactions to Cas9, and challenges associated with delivery methods. Despite these limitations, this review aims to provide knowledge about CRISPR/Cas9 genome editing tools and explore how they can help overcome resistance in patients with respiratory disorders. Additionally, this study discusses concerns related to applications of CRISPR and provides an overview of successful clinical trial studies.

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Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. We show here that in a subset of these systems, the mature crRNA that is base-paired to trans-activating crRNA (tracrRNA) forms a two-RNA structure that directs the CRISPR-associated protein Cas9 to introduce double-stranded (ds) breaks in target DNA. At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand, whereas the Cas9 RuvC-like domain cleaves the noncomplementary strand. The dual-tracrRNA:crRNA, when engineered as a single RNA chimera, also directs sequence-specific Cas9 dsDNA cleavage. Our study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.

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CRISPR-Cas9-based genetic screens are a powerful new tool in biology. By simply altering the sequence of the single-guide RNA (sgRNA), Cas9 can be reprogrammed to target different sites in the genome with relative ease, but the on-target activity and off-target effects of individual sgRNAs can vary widely. Here, we use recently-devised sgRNA design rules to create human and mouse genome-wide libraries, perform positive and negative selection screens and observe that the use of these rules produced improved results. Additionally, we profile the off-target activity of thousands of sgRNAs and develop a metric to predict off-target sites. We incorporate these findings from large-scale, empirical data to improve our computational design rules and create optimized sgRNA libraries that maximize on-target activity and minimize off-target effects to enable more effective and efficient genetic screens and genome engineering.

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Author and article information

Contributors

Soudeh Ghafouri-Fard: s.ghafourifard@sbmu.ac.ir

Mohammad Taheri: Mohammad.taheri@uni-jena.de

Journal

Journal ID (nlm-ta): Cell Commun Signal

Journal ID (iso-abbrev): Cell Commun Signal

Title: Cell Communication and Signaling : CCS

Publisher: BioMed Central (London )

ISSN (Electronic): 1478-811X

Publication date (Electronic): 14 June 2024

Publication date PMC-release: 14 June 2024

Publication date Collection: 2024

Volume: 22

Electronic Location Identifier: 329

Affiliations

[1 ]Department of Biomedical Sciences, College of Science, Cihan University-Erbil, ( https://ror.org/03hevjm30) Erbil, 44001 Kurdistan Region Iraq

[2 ]Department of Clinical Analysis, College of Pharmacy, Hawler Medical University, ( https://ror.org/02a6g3h39) Kurdistan Region, Erbil, Iraq

[3 ]Research Center, University of Halabja, ( https://ror.org/037fm3958) Halabja, 46018 Kurdistan region Iraq

[4 ]Medical Laboratory Science, College of Health Sciences, Lebanese French University, ( https://ror.org/030t96b35) Kurdistan Region, Erbil, Iraq

[5 ]Department of Pharmaceutical Basic Science, Tishk International University, ( https://ror.org/03pbhyy22) Kurdistan Region, Iraq

[6 ]General Directorate of Scientific Research Center, Salahaddin University-Erbil, ( https://ror.org/02124dd11) Erbil, Kurdistan Region Iraq

[7 ]Department of Medical Genetics, Shahid Beheshti University of Medical Sciences, ( https://ror.org/034m2b326) Tehran, Iran

[8 ]Institute of Human Genetics, Jena University Hospital, ( https://ror.org/035rzkx15) Jena, Germany

Article

Publisher ID: 1713

DOI: 10.1186/s12964-024-01713-8

PMC ID: 11179281

PubMed ID: 38877530

SO-VID: 1f73516c-adc4-4302-a23d-94dd6f8020fd

License:

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

History

Date received : 5 March 2024

Date accepted : 12 June 2024

Funding

Funded by: Universitätsklinikum Jena (8979)

Open Access :

Open Access funding enabled and organized by Projekt DEAL.

Custom metadata

ScienceOpen disciplines: Cell biology

Keywords: respiratory disease,crispr/ cas9,drug resistance

Data availability:

ScienceOpen disciplines: Cell biology

Keywords: respiratory disease, crispr/ cas9, drug resistance

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