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CRISPR/Cas9: From the Bacteria to Widespread Advanced Genome Editing Tools...
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CRISPR/Cas9: From the Bacteria to Widespread Advanced Genome Editing Tools in Mammalian Cells

A special issue of Cells (ISSN 2073-4409). This special issue belongs to the section "Cell Methods".

Deadline for manuscript submissions: closed (31 January 2024) | Viewed by 7105

Special Issue Editors

Dr. Sergiu Chira

E-Mail Website
Guest Editor
Research Center for Functional Genomics, Biomedicine and Translational Medicine, "Iuliu Hatieganu" University of Medicine and Pharmacy, 400337 Cluj-Napoca, Romania
Interests: molecular biology; cell biology; gene therapy; genome engineering; non-viral vectors; viral vectors
Prof. Dr. Yusuke Kamachi

E-Mail Website
Co-Guest Editor
School of Environmental Science and Engineering, Kochi University of Technology, Kami, Kochi 782-8502, Japan
Interests: developmental biology; zebrafish; genome editing; CRISPR; gene tagging; cell differentiation; cell-type specification; transcriptional regulation; Sox transcription factors; stem cells
Dr. Cecilia Bica

E-Mail Website
Co-Guest Editor
Research Center for Functional Genomics, Biomedicine and Translational Medicine, "Iuliu Hatieganu" University of Medicine and Pharmacy, 400337 Cluj-Napoca, Romania
Interests: bioinfromatics; cancer; cancer biomarkers; tumor biology; tumor microenvironment

Special Issue Information

Dear Colleagues,

Microbiology has been probably one of the most important sources of inspiration for scientists that led to the development of numerous fields of research, such as molecular biology, biotechnology, and biomedical sciences. Initially described as a defense mechanism of bacteria against invading phage viruses, CRISPR/Cas9 is now a leading tool for editing and regulation of gene function in mammalian cell’s genome, surpassing other gene editing technologies, such as TALEN and ZFN, in terms of versatility and ease of targeting virtually any genomic locus. This innovating technology holds the promise of achieving precise modifications in the genome, broadening the application fields of CRISPR/Cas9 from gene functional studies to disease models and therapeutic concepts. Implementing such technology for treatment of human diseases, and, in particular, for malignant disorders, is a goal for which great efforts have been made in recent years. However, few studies have reached clinical trials, and are mainly restricted to ex vivo gene edited cells for immunotherapies, and to some extent as a pathogen detection tool. These shortcomings relate to “off-target” cleavage of genomic loci, an event that might result in undesired side effects, and even promoting a malignant behavior of the CRISPR/Cas9 edited cells. In this regard, novel variants of CRISPR/Cas9 with improved specificity and novel functions have been developed, in addition to evaluating delivery strategies for safe and efficient targeting of the diseased cells.

The current Special Issue will accept original studies, reviews, and technical reports in the field of CRISPR/Cas9 research meant to cover the spectrum of diverse applications, both of fundamental and translational value.

Dr. Sergiu Chira
Prof. Dr. Yusuke Kamachi
Dr. Cecilia Bica
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Cells is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • bacteria
  • CRISPR/Cas9
  • genome editing
  • gene regulation
  • delivery
  • targeted therapy

Published Papers (2 papers)

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Research

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11 pages, 2526 KiB  
Communication
PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies
by Guoliang Yuan, Stanton Martin, Md Mahmudul Hassan, Gerald A. Tuskan and Xiaohan Yang
Cells 2022, 11(16), 2467; https://doi.org/10.3390/cells11162467 - 09 Aug 2022
Cited by 8 | Viewed by 2934
Abstract
Multiplexed CRISPR technologies have great potential for pathway engineering and genome editing. However, their applications are constrained by complex, laborious and time-consuming cloning steps. In this research, we developed a novel method, PARA, which allows for the one-step assembly of multiple guide RNAs [...] Read more.
Multiplexed CRISPR technologies have great potential for pathway engineering and genome editing. However, their applications are constrained by complex, laborious and time-consuming cloning steps. In this research, we developed a novel method, PARA, which allows for the one-step assembly of multiple guide RNAs (gRNAs) into a CRISPR vector with up to 18 gRNAs. Here, we demonstrate that PARA is capable of the efficient assembly of transfer RNA/Csy4/ribozyme-based gRNA arrays. To aid in this process and to streamline vector construction, we developed a user-friendly PARAweb tool for designing PCR primers and component DNA parts and simulating assembled gRNA arrays and vector sequences. Full article
(This article belongs to the Special Issue CRISPR/Cas9: From the Bacteria to Widespread Advanced Genome Editing Tools in Mammalian Cells)
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Review

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20 pages, 7180 KiB  
Review
Genome Editing Approaches with CRISPR/Cas9 for Cancer Treatment: Critical Appraisal of Preclinical and Clinical Utility, Challenges, and Future Research
by Sergiu Chira, Andreea Nutu, Ecaterina Isacescu, Cecilia Bica, Laura Pop, Cristina Ciocan and Ioana Berindan-Neagoe
Cells 2022, 11(18), 2781; https://doi.org/10.3390/cells11182781 - 06 Sep 2022
Cited by 5 | Viewed by 3694
Abstract
The increasing burden on human malignant diseases became a major concern for healthcare practitioners, that must deal with tumor relapse and the inability to efficiently treat metastasis, in addition to side effects. Throughout the decades, many therapeutic strategies have been employed to improve [...] Read more.
The increasing burden on human malignant diseases became a major concern for healthcare practitioners, that must deal with tumor relapse and the inability to efficiently treat metastasis, in addition to side effects. Throughout the decades, many therapeutic strategies have been employed to improve the clinical outcomes of cancer patients and great efforts have been made to develop more efficient and targeted medicines. The malignant cell is characterized by genetic and epigenetic modifications, therefore targeting those specific drivers of carcinogenesis is highly desirable. Among the genome editing technologies, CRISPR/Cas9 stood as a promising candidate for cancer treatment alternatives, due to its low complexity design. First described as a defense mechanism of bacteria against invading foreign DNA, later it was shown that CRISPR components can be engineered to target specific DNA sequences in a test tube, a discovery that was awarded later with the Nobel Prize in chemistry for its rapid expansion as a reliable genome editing tool in many fields of research, including medicine. The present paper aims of describing CRISPR/Cas9 potential targets for malignant disorders, and the approaches used for achieving this goal. Aside from preclinical studies, we also present the clinical trials that use CRISPR-based technology for therapeutic purposes of cancer. Finally, a summary of the presented studies adds a more focused view of the therapeutic value CRISPR/Cas9 holds and the associated shortcomings. Full article
(This article belongs to the Special Issue CRISPR/Cas9: From the Bacteria to Widespread Advanced Genome Editing Tools in Mammalian Cells)
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