scholarly journals The Missing Link: Cre Pigs for Cancer Research

2021 ◽  
Vol 11 ◽  
Author(s):  
Daniela Kalla ◽  
Krzysztof Flisikowski ◽  
Kaiyuan Yang ◽  
Laura Beltran Sangüesa ◽  
Mayuko Kurome ◽  
...  
Keyword(s):  
Cancer Research ◽  
Basic Research ◽  
Human Pancreatic Cancer ◽  
Specific Expression ◽  
Fluorescent Reporter ◽  
Temporal Gene Expression ◽  
Human Cancers ◽  
Complementary Resources ◽  
Cancer Models ◽  
Large Animals

The Cre/loxP system is a powerful tool for the generation of animal models with precise spatial and temporal gene expression. It has proven indispensable in the generation of cancer models with tissue specific expression of oncogenes or the inactivation of tumor suppressor genes. Consequently, Cre-transgenic mice have become an essential prerequisite in basic cancer research. While it is unlikely that pigs will ever replace mice in basic research they are already providing powerful complementary resources for translational studies. But, although conditionally targeted onco-pigs have been generated, no Cre-driver lines exist for any of the major human cancers. To model human pancreatic cancer in pigs, Cre-driver lines were generated by CRISPR/Cas9-mediated insertion of codon-improved Cre (iCre) into the porcine PTF1A gene, thus guaranteeing tissue and cell type specific function which was proven using dual fluorescent reporter pigs. The method used can easily be adapted for the generation of other porcine Cre-driver lines, providing a missing tool for modeling human cancers in large animals.

Applications and advances of CRISPR/Cas9 in animal cancer model

2020 ◽  
Vol 19 (3) ◽  
pp. 235-241 ◽  
Author(s):  
Min Xu ◽  
Qiaoyou Weng ◽  
Jiansong Ji
Keyword(s):  
Cancer Research ◽  
Gene Editing ◽  
Cancer Development ◽  
Cancer Model ◽  
Associate Protein ◽  
Human Cancers ◽  
Cancer Models ◽  
Multistep Process ◽  
Recent Developments

Abstract The recent developments of clustered regularly interspaced short palindromic repeats(CRISPR)/-associate protein 9 (CRISPR/Cas9) have got scientific interests due to the straightforward, efficient and versatile talents of it. Furthermore, the CRISPR/Cas9 system has democratized access to gene editing in many biological fields, including cancer. Cancer development is a multistep process caused by innate and acquired mutations and leads to the initiation and progression of tumorigenesis. It is obvious that establishing appropriate animal cancer models which can simulate human cancers is crucial for cancer research currently. Since the emergence of CRISPR/Cas9, considerable efforts have been taken by researchers to apply this technology in generating animal cancer models. Although there is still a long way to go we are happy to see the achievements we have made and the promising future we have.

scholarly journals Genetically Engineered Pigs to Study Cancer

2020 ◽  
Vol 21 (2) ◽  
pp. 488 ◽  
Author(s):  
Daniela Kalla ◽  
Alexander Kind ◽  
Angelika Schnieke
Keyword(s):  
Cancer Research ◽  
Animal Models ◽  
Basic Research ◽  
Genetically Engineered ◽  
Complementary Information ◽  
Oncogenic Mutation ◽  
Cancer Models ◽  
Therapeutic Procedures ◽  
Underlying Mechanisms ◽  
Near Future

Recent decades have seen groundbreaking advances in cancer research. Genetically engineered animal models, mainly in mice, have contributed to a better understanding of the underlying mechanisms involved in cancer. However, mice are not ideal for translating basic research into studies closer to the clinic. There is a need for complementary information provided by non-rodent species. Pigs are well suited for translational biomedical research as they share many similarities with humans such as body and organ size, aspects of anatomy, physiology and pathophysiology and can provide valuable means of developing and testing novel diagnostic and therapeutic procedures. Porcine oncology is a new field, but it is clear that replication of key oncogenic mutation in pigs can usefully mimic several human cancers. This review briefly outlines the technology used to generate genetically modified pigs, provides an overview of existing cancer models, their applications and how the field may develop in the near future.

scholarly journals GYNOCARE Update: Modern Strategies to Improve Diagnosis and Treatment of Rare Gynecologic Tumors—Current Challenges and Future Directions

Cancers ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 493
Author(s):  
Riccardo Di Fiore ◽  
Sherif Suleiman ◽  
Bridget Ellul ◽  
Sharon A. O’Toole ◽  
Charles Savona-Ventura ◽  
...  
Keyword(s):  
Cancer Research ◽  
Gynecological Cancer ◽  
Gynecologic Cancer ◽  
Delayed Diagnosis ◽  
Research Programme ◽  
Basic Research ◽  
Diagnosis And Treatment ◽  
Rare Cancer ◽  
Cost Action ◽  
Gynecologic Tumors

More than 50% of all gynecologic tumors can be classified as rare (defined as an incidence of ≤6 per 100,000 women) and usually have a poor prognosis owing to delayed diagnosis and treatment. In contrast to almost all other common solid tumors, the treatment of rare gynecologic tumors (RGT) is often based on expert opinion, retrospective studies, or extrapolation from other tumor sites with similar histology, leading to difficulty in developing guidelines for clinical practice. Currently, gynecologic cancer research, due to distinct scientific and technological challenges, is lagging behind. Moreover, the overall efforts for addressing these challenges are fragmented across different European countries and indeed, worldwide. The GYNOCARE, COST Action CA18117 (European Network for Gynecological Rare Cancer Research) programme aims to address these challenges through the creation of a unique network between key stakeholders covering distinct domains from concept to cure: basic research on RGT, biobanking, bridging with industry, and setting up the legal and regulatory requirements for international innovative clinical trials. On this basis, members of this COST Action, (Working Group 1, “Basic and Translational Research on Rare Gynecological Cancer”) have decided to focus their future efforts on the development of new approaches to improve the diagnosis and treatment of RGT. Here, we provide a brief overview of the current state-of-the-art and describe the goals of this COST Action and its future challenges with the aim to stimulate discussion and promote synergy across scientists engaged in the fight against this rare cancer worldwide.

scholarly journals A Method for Rapid Demineralization of Teeth and Bones

2010 ◽  
Vol 4 (1) ◽  
pp. 223-229 ◽  
Author(s):  
Andrew Cho ◽  
Shigeki Suzuki ◽  
Junko Hatakeyama ◽  
Naoto Haruyama ◽  
Ashok B Kulkarni
Keyword(s):  
Reporter Gene ◽  
Gene Expression Pattern ◽  
Cell Lineage ◽  
Careful Analysis ◽  
Lacz Gene ◽  
Bone Specimen ◽  
Specific Expression ◽  
Temporal Gene Expression ◽  
Gene And Protein Expression ◽  
Tissue Specimens

Tooth and bone specimen require extensive demineralization for careful analysis of cell morphology, as well as gene and protein expression levels. The LacZ gene, which encodes the ß-galactosidase enzyme, is often used as a reporter gene to study gene-structure function, tissue-specific expression by a promoter, cell lineage and fate. This reporter gene is particularly useful for analyzing the spatial and temporal gene expression pattern, by expressing the LacZ gene under the control of a promoter of interest. To analyze LacZ activity, and the expression of other genes and their protein products in teeth and bones, it is necessary to carry out a complete demineralization of the specimen before cutting sections. However, strong acids, such as formic acid used for tooth demineralization, destroy the activities of enzymes including those of ß-galactosidase. Therefore, most protocols currently use mild acids such as 0.1 M ethylene diamine tetra-acetic acid (EDTA) for demineralization of tooth and bone specimen, which require a longer period of treatment for complete demineralization. A method by which hard tissue specimens such as teeth and bones can be rapidly, but gently, decalcified is necessary to save time and effort. Here, we report a suitable method for rapid demineralization of mouse teeth in 0.1M EDTA at 42˚C without any loss of ß-galactosidase activity.

Proteomics in cancer research

2002 ◽  
Vol 12 (5) ◽  
pp. 409-423
Author(s):  
W. Wu ◽  
W. Hu ◽  
J. J. Kavanagh
Keyword(s):  
Cancer Research ◽  
Drug Targets ◽  
Review Paper ◽  
Basic Research ◽  
Anticancer Therapy ◽  
Molecular Medicine ◽  
Clinical Cancer Research ◽  
Detection And Diagnosis ◽  
Early Detection And Diagnosis ◽  
Novel Protein

With the human genome sequence now determined, the field of molecular medicine is moving beyond genomics to proteomics. In the field of cancer research, the key question is: how can oncologists best use techniques of proteomics in basic research and clinical application? In the postgenomic era, proteomics promises the discovery of biomarkers and tumor markers for early detection and diagnosis, novel protein-based drug targets for anticancer therapy, and new endpoints for the assessment of therapeutic efficacy and toxicity. This review paper will explore key themes in proteomics and their application in clinical cancer research.

scholarly journals Genetic Engineering of Zebrafish in Cancer Research

Cancers ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 2168
Author(s):  
Ludivine Raby ◽  
Pamela Völkel ◽  
Xuefen Le Bourhis ◽  
Pierre-Olivier Angrand
Keyword(s):  
Genetic Alterations ◽  
Therapeutic Strategies ◽  
Molecular Signatures ◽  
Reverse Genetic ◽  
Human Cancers ◽  
Oncogenic Mutations ◽  
Cancer Models ◽  
Conditional Expression ◽  
Cancer Initiation

Zebrafish (Danio rerio) is an excellent model to study a wide diversity of human cancers. In this review, we provide an overview of the genetic and reverse genetic toolbox allowing the generation of zebrafish lines that develop tumors. The large spectrum of genetic tools enables the engineering of zebrafish lines harboring precise genetic alterations found in human patients, the generation of zebrafish carrying somatic or germline inheritable mutations or zebrafish showing conditional expression of the oncogenic mutations. Comparative transcriptomics demonstrate that many of the zebrafish tumors share molecular signatures similar to those found in human cancers. Thus, zebrafish cancer models provide a unique in vivo platform to investigate cancer initiation and progression at the molecular and cellular levels, to identify novel genes involved in tumorigenesis as well as to contemplate new therapeutic strategies.

scholarly journals Application of the CRISPR/Cas9-based gene editing technique in basic research, diagnosis, and therapy of cancer

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Huimin Zhang ◽  
Chunhong Qin ◽  
Changming An ◽  
Xiwang Zheng ◽  
Shuxin Wen ◽  
...  
Keyword(s):  
Cancer Research ◽  
Gene Editing ◽  
Basic Research ◽  
Genomic Locus ◽  
Future Directions ◽  
Genomic Changes ◽  
Diagnosis And Therapy ◽  
Oncology Research ◽  
Potential Applications ◽  
A Site

AbstractThe 2020 Nobel Prize in Chemistry was awarded to Emmanuelle Charpentier and Jennifer Doudna for the development of the Clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease9 (CRISPR/Cas9) gene editing technology that provided new tools for precise gene editing. It is possible to target any genomic locus virtually using only a complex nuclease protein with short RNA as a site-specific endonuclease. Since cancer is caused by genomic changes in tumor cells, CRISPR/Cas9 can be used in the field of cancer research to edit genomes for exploration of the mechanisms of tumorigenesis and development. In recent years, the CRISPR/Cas9 system has been increasingly used in cancer research and treatment and remarkable results have been achieved. In this review, we introduced the mechanism and development of the CRISPR/Cas9-based gene editing system. Furthermore, we summarized current applications of this technique for basic research, diagnosis and therapy of cancer. Moreover, the potential applications of CRISPR/Cas9 in new emerging hotspots of oncology research were discussed, and the challenges and future directions were highlighted.

scholarly journals High-fidelity Cas13 variants for targeted RNA degradation with minimal collateral effect

2021 ◽  
Author(s):  
Huawei Tong ◽  
Jia Huang ◽  
Qingquan Xiao ◽  
Bingbing He ◽  
Xue Dong ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Basic Research ◽  
Rna Degradation ◽  
High Fidelity ◽  
Reporter System ◽  
Fluorescent Reporter ◽  
Major Barrier ◽  
Collateral Effects ◽  
Collateral Effect

CRISPR-Cas13 systems have recently been employed for targeted RNA degradation in various organisms. However, collateral degradation of bystander RNAs has imposed a major barrier for their in vivo applications. We designed a dual-fluorescent reporter system for detecting collateral effects and screening Cas13 variants in mammalian cells. Among over 200 engineered variants, several Cas13 variants (including Cas13d and Cas13X) exhibit efficient on-target activity but markedly reduced collateral activity. Furthermore, transcriptome-wide off-targets and cell growth arrest induced by Cas13 are absent for these variants. Importantly, high-fidelity Cas13 variants show comparable RNA knockdown activity with wild-type Cas13 but no detectable collateral damage in transgenic mice and adeno-associated virus-mediated somatic cell targeting. Thus, high-fidelity Cas13 variants with minimal collateral effect are now available for targeted degradation of RNAs in basic research and therapeutic applications.

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