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Cancers
Special Issues
CRISPR-Mediated Cancer Modeling
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CRISPR-Mediated Cancer Modeling

A special issue of Cancers (ISSN 2072-6694). This special issue belongs to the section "Methods and Technologies Development".

Deadline for manuscript submissions: closed (31 May 2022) | Viewed by 11980

Special Issue Editor

Dr. Martin Kristian Thomsen

E-Mail Website
Guest Editor
Department of Biomedicine, Aarhus University, 8000 Aarhus, Denmark
Interests: prostate cancer; GBM; lung cancer; mouse models; CRISPR; AAV; multiplexing CRISPR guides

Special Issue Information

Dear Colleagues,

The discovery of the CRISPR/Cas System was rewarded with the Nobel Prize in 2020, recipients being researchers Emmanuelle Charpentier and Jennifer Doudna. CRISPR technology has revolutionized many areas of research, including cancer research. The possibility to easily generate loss of function studies by the application of CRISPR has opened new possibilities of research. In addition, gain of function studies can be conducted by homologues repaired after CRISPR breaks. This has allowed research to model common loss or gain of function mutations that occur in cancer. CRISPR technology is now used for genome-wide screening by the application of large libraries as well as temporal gene activation or inhibition. Furthermore, as these technologies are both applied in vitro and in vivo, this has opened new research avenues regarding the complexity of cancer biology.

This Special Issue will focus on cancer research where the application of CRISPR has been applied. We will aim to publish articles in this issue where the authors’ application of technology has provided new insight into cancer biology. This issue should contain studies from in vitro, ex vivo, and in vivo applications in order to provide a full understanding of the possibilities of applying CRISPR technology in cancer research.

Dr. Martin Kristian Thomsen
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 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. Cancers 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 2900 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

  • cancer
  • CRISPR
  • CRISPRi
  • CRISPRa
  • AAV
  • CRISPR libraries
  • in vivo
  • in vitro
  • CRISPR KO

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Published Papers (3 papers)

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Research

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13 pages, 3921 KiB  
Article
In Vivo Application of CRISPR/Cas9 Revealed Implication of Foxa1 and Foxp1 in Prostate Cancer Proliferation and Epithelial Plasticity
by Huiqiang Cai, Simon N. Agersnap, Amalie Sjøgren, Mikkel K. Simonsen, Mathilde S. Blaavand, Ulrikke V. Jensen and Martin K. Thomsen
Cancers 2022, 14(18), 4381; https://doi.org/10.3390/cancers14184381 - 8 Sep 2022
Cited by 11 | Viewed by 2883
Abstract
Prostate cancer is the most common cancer in men in the Western world and the number is rising. Prostate cancer is notoriously heterogeneous, which makes it hard to generate and study in pre-clinical models. The family of Forkhead box (FOX) transcription factors are [...] Read more.
Prostate cancer is the most common cancer in men in the Western world and the number is rising. Prostate cancer is notoriously heterogeneous, which makes it hard to generate and study in pre-clinical models. The family of Forkhead box (FOX) transcription factors are often altered in prostate cancer with especially high mutation burden in FOXA1 and FOXP1. FOXA1 harbors loss or gain of function mutations in 8% of prostate cancer, which increases to 14% in metastatic samples. FOXP1 predominately occurs with loss of function mutations in 7% of primary tumors, and similar incidents are found in metastatic samples. Here, we applied in vivo CRISPR editing, to study the loss of functions of these two FOX transcription factors, in murine prostate in combination with loss of Pten. Deficiency of Foxp1 increased proliferation in combination with loss of Pten. In contrast, proliferation was unchanged when androgen was deprived. The expression of Tmprss2 was increased when Foxp1 was mutated in vivo, showing that Foxp1 is a repressor for this androgen-regulated target. Furthermore, analysis of FOXP1 and TMPRSS2 expression in a human prostate cancer data set revealed a negative correlation. Mutation of Foxa1 in the murine prostate induces cell plasticity to luminal cells. Here, epithelial cells with loss of Foxa1 were transdifferentiated to cells with expression of the basal markers Ck5 and p63. Interestingly, these cells were located in the lumen and did not co-express Ck8. Overall, this study reveals that loss of Foxp1 increases cell proliferation, whereas loss of Foxa1 induces epithelial plasticity in prostate cancer. Full article
(This article belongs to the Special Issue CRISPR-Mediated Cancer Modeling)
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17 pages, 3181 KiB  
Article
The CRISPR/Cas9 Minipig—A Transgenic Minipig to Produce Specific Mutations in Designated Tissues
by Martin Fogtmann Berthelsen, Maria Riedel, Huiqiang Cai, Søren H. Skaarup, Aage K. O. Alstrup, Frederik Dagnæs-Hansen, Yonglun Luo, Uffe B. Jensen, Henrik Hager, Ying Liu, Henrik Callesen, Mikkel H. Vendelbo, Jannik E. Jakobsen and Martin Kristian Thomsen
Cancers 2021, 13(12), 3024; https://doi.org/10.3390/cancers13123024 - 16 Jun 2021
Cited by 12 | Viewed by 4974
Abstract
The generation of large transgenic animals is impeded by complex cloning, long maturation and gastrulation times. An introduction of multiple gene alterations increases the complexity. We have cloned a transgenic Cas9 minipig to introduce multiple mutations by CRISPR in somatic cells. Transgenic Cas9 [...] Read more.
The generation of large transgenic animals is impeded by complex cloning, long maturation and gastrulation times. An introduction of multiple gene alterations increases the complexity. We have cloned a transgenic Cas9 minipig to introduce multiple mutations by CRISPR in somatic cells. Transgenic Cas9 pigs were generated by somatic cell nuclear transfer and were backcrossed to Göttingen Minipigs for two generations. Cas9 expression was controlled by FlpO-mediated recombination and was visualized by translation from red to yellow fluorescent protein. In vitro analyses in primary fibroblasts, keratinocytes and lung epithelial cells confirmed the genetic alterations executed by the viral delivery of single guide RNAs (sgRNA) to the target cells. Moreover, multiple gene alterations could be introduced simultaneously in a cell by viral delivery of sgRNAs. Cells with loss of TP53, PTEN and gain-of-function mutation in KRASG12D showed increased proliferation, confirming a transformation of the primary cells. An in vivo activation of Cas9 expression could be induced by viral delivery to the skin. Overall, we have generated a minipig with conditional expression of Cas9, where multiple gene alterations can be introduced to somatic cells by viral delivery of sgRNA. The development of a transgenic Cas9 minipig facilitates the creation of complex pre-clinical models for cancer research. Full article
(This article belongs to the Special Issue CRISPR-Mediated Cancer Modeling)
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Review

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12 pages, 1338 KiB  
Review
Application of CRISPR for In Vivo Mouse Cancer Studies
by Martin K. Thomsen
Cancers 2022, 14(20), 5014; https://doi.org/10.3390/cancers14205014 - 13 Oct 2022
Cited by 6 | Viewed by 3487
Abstract
Clustered regularly interspaced short palindromic repeats (CRISPR) are widely used in cancer research to edit specific genes and study their functions. This applies both to in vitro and in vivo studies where CRISPR technology has accelerated the generation of specific loss- or gain-of-function [...] Read more.
Clustered regularly interspaced short palindromic repeats (CRISPR) are widely used in cancer research to edit specific genes and study their functions. This applies both to in vitro and in vivo studies where CRISPR technology has accelerated the generation of specific loss- or gain-of-function mutations. This review focuses on CRISPR for generating in vivo models of cancer by editing somatic cells in specific organs. The delivery of CRISPR/Cas to designated tissues and specific cell compartments is discussed with a focus on different methods and their advantages. One advantage of CRISPR/Cas is the possibility to target multiple genes simultaneously in the same cell and therefore generate complex mutation profiles. This complexity challenges the interpretation of results and different methods to analyze the samples discussed herein. CRISPR-induced tumors are also different from classical tumors in pre-clinical models. Especially the clonal evolution of CRISPR-induced tumors adds new insight into cancer biology. Finally, the review discusses future perspectives for CRISPR technology in pre-clinical models with a focus on in vivo screening, CRISPR activation/inhibition, and the development of prime/ base-editing for the introduction of specific gene editing. Full article
(This article belongs to the Special Issue CRISPR-Mediated Cancer Modeling)
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