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Cells
Special Issues
CRISPR-Based Genome Editing Approaches in Cancer Therapy
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CRISPR-Based Genome Editing Approaches in Cancer Therapy

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

Deadline for manuscript submissions: 30 November 2025 | Viewed by 975

Special Issue Editor

Dr. Martin Pal

E-Mail Website
Guest Editor
School of Dentistry and Medical Sciences, Charles Sturt University, Wagga Wagga, Australia
Interests: genome editing; CRISPR-technology

Special Issue Information

Dear Colleagues,

This Special Issue explores the role of CRISPR-based genome editing approaches in cancer therapy. Genome editing using RNA-guided programmable clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nucleases has revolutionized genetic research, offering unparalleled speed, efficiency, and precision. Since its first application in mammalian cells more than a decade ago, the CRISPR toolbox has rapidly expanded, continuously offering novel genetic tools aiding disease modeling and therapeutic approaches, including in cancer.

For this Special Issue, we welcome submissions that harness novel advancements of CRISPR-technology aimed at improving cancer treatments, including but not limited to CRISPR-based genome editing in cancer cells, immunotherapy approaches using CRISPR technology, CRISPR diagnostics for early cancer detection and monitoring, or the use of functional genomic screens to identify novel cancer-driving genes. Studies highlighting the role of using CRISPR-based genome editing for anti-cancer treatments are of particular interest.

By highlighting innovative research and translational applications, we endeavor to advance our understanding of how CRISPR-based genome editing approaches can shape personalized anti-cancer treatments.

Dr. Martin Pal
Guest Editor

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

  • CRISPR
  • genome editing
  • cancer therapy
  • precision medicine

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Further information on MDPI's Special Issue policies can be found here.

Published Papers (1 paper)

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Research

20 pages, 5588 KiB  
Article
Rapid and Robust Generation of Homozygous Fluorescent Reporter Knock-In Cell Pools by CRISPR-Cas9
by Jicheng Yang, Fusheng Guo, Hui San Chin, Gao Bin Chen, Ziyan Zhang, Lewis Williams, Andrew J. Kueh, Pierce K. H. Chow, Marco J. Herold and Nai Yang Fu
Cells 2025, 14(15), 1165; https://doi.org/10.3390/cells14151165 - 29 Jul 2025
Viewed by 673
Abstract
Conventional methods for generating knock-out or knock-in mammalian cell models using CRISPR-Cas9 genome editing often require tedious single-cell clone selection and expansion. In this study, we develop and optimise rapid and robust strategies to engineer homozygous fluorescent reporter knock-in cell pools with precise [...] Read more.
Conventional methods for generating knock-out or knock-in mammalian cell models using CRISPR-Cas9 genome editing often require tedious single-cell clone selection and expansion. In this study, we develop and optimise rapid and robust strategies to engineer homozygous fluorescent reporter knock-in cell pools with precise genome editing, circumventing clonal variability inherent to traditional approaches. To reduce false-positive cells associated with random integration, we optimise the design of donor DNA by removing the start codon of the fluorescent reporter and incorporating a self-cleaving T2A peptide system. Using fluorescence-assisted cell sorting (FACS), we efficiently identify and isolate the desired homozygous fluorescent knock-in clones, establishing stable cell pools that preserve parental cell line heterogeneity and faithfully reflect endogenous transcriptional regulation of the target gene. We evaluate the knock-in efficiency and rate of undesired random integration in the electroporation method with either a dual-plasmid system (sgRNA and donor DNA in two separate vectors) or a single-plasmid system (sgRNA and donor DNA combined in one vector). We further demonstrate that coupling our single-plasmid construct with an integrase-deficient lentivirus vector (IDLV) packaging system efficiently generates fluorescent knock-in reporter cell pools, offering flexibility between electroporation and lentivirus transduction methods. Notably, compared to the electroporation methods, the IDLV system significantly minimises random integration. Moreover, the resulting reporter cell lines are compatible with most of the available genome-wide sgRNA libraries, enabling unbiased CRISPR screens to identify key transcriptional regulators of a gene of interest. Overall, our methodologies provide a powerful genetic tool for rapid and robust generation of fluorescent reporter knock-in cell pools with precise genome editing by CRISPR-Cas9 for various research purposes. Full article
(This article belongs to the Special Issue CRISPR-Based Genome Editing Approaches in Cancer Therapy)
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