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 Conditional Reprogramming for Patient-Derived Cancer Models and Next-Generation Living Biobanks

Cells ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 1327 ◽  
Author(s):  
Palechor-Ceron ◽  
Krawczyk ◽  
Dakic ◽  
Simic ◽  
Yuan ◽  
...  
Keyword(s):  
Cancer Research ◽  
Cancer Biology ◽  
Tumor Tissue ◽  
Precision Oncology ◽  
Next Generation ◽  
Cancer Model ◽  
Functional Diagnostics ◽  
Differentiation Potential ◽  
Cell Lung Carcinoma ◽  
Cancer Models

Traditional cancer models including cell lines and animal models have limited applications in both basic and clinical cancer research. Genomics-based precision oncology only help 2–20% patients with solid cancer. Functional diagnostics and patient-derived cancer models are needed for precision cancer biology. In this review, we will summarize applications of conditional cell reprogramming (CR) in cancer research and next generation living biobanks (NGLB). Together with organoids, CR has been cited in two NCI (National Cancer Institute, USA) programs (PDMR: patient-derived cancer model repository; HCMI: human cancer model initiatives. HCMI will be distributed through ATCC). Briefly, the CR method is a simple co-culture technology with a Rho kinase inhibitor, Y-27632, in combination with fibroblast feeder cells, which allows us to rapidly expand both normal and malignant epithelial cells from diverse anatomic sites and mammalian species and does not require transfection with exogenous viral or cellular genes. Establishment of CR cells from both normal and tumor tissue is highly efficient. The robust nature of the technique is exemplified by the ability to produce 2 × 106 cells in five days from a core biopsy of tumor tissue. Normal CR cell cultures retain a normal karyotype and differentiation potential and CR cells derived from tumors retain their tumorigenic phenotype. CR also allows us to enrich cancer cells from urine (for bladder cancer), blood (for prostate cancer), and pleural effusion (for non-small cell lung carcinoma). The ability to produce inexhaustible cell populations using CR technology from small biopsies and cryopreserved specimens has the potential to transform biobanking repositories (NGLB: next-generation living biobank) and current pathology practice by enabling genetic, biochemical, metabolomic, proteomic, and biological assays, including chemosensitivity testing as a functional diagnostics tool for precision cancer medicine. We discussed analyses of patient-derived matched normal and tumor models using a case with tongue squamous cell carcinoma as an example. Last, we summarized applications in cancer research, disease modeling, drug discovery, and regenerative medicine of CR-based NGLB.

scholarly journals Review of applications of CRISPR-Cas9 gene-editing technology in cancer research

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Ziyi Zhao ◽  
Chenxi Li ◽  
Fei Tong ◽  
Jingkuang Deng ◽  
Guofu Huang ◽  
...  
Keyword(s):  
Cancer Research ◽  
Cancer Progression ◽  
Tumor Suppressors ◽  
Genetic Screening ◽  
Gene Editing ◽  
Causes Of Death ◽  
Cancer Models ◽  
Cancer Genomes ◽  
Passenger Mutations ◽  
Cancer Studies

AbstractCharacterized by multiple complex mutations, including activation by oncogenes and inhibition by tumor suppressors, cancer is one of the leading causes of death. Application of CRISPR-Cas9 gene-editing technology in cancer research has aroused great interest, promoting the exploration of the molecular mechanism of cancer progression and development of precise therapy. CRISPR-Cas9 gene-editing technology provides a solid basis for identifying driver and passenger mutations in cancer genomes, which is of great value in genetic screening and for developing cancer models and treatments. This article reviews the current applications of CRISPR-Cas9 gene-editing technology in various cancer studies, the challenges faced, and the existing solutions, highlighting the potential of this technology for cancer treatment.

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.

scholarly journals Advances in Cereal Crop Genomics for Resilience under Climate Change

Life ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 502
Author(s):  
Tinashe Zenda ◽  
Songtao Liu ◽  
Anyi Dong ◽  
Huijun Duan
Keyword(s):  
Climate Change ◽  
Genome Sequencing ◽  
Gene Editing ◽  
Agronomic Traits ◽  
Cereal Crops ◽  
Agricultural System ◽  
Plant Genomics ◽  
Crop Species ◽  
Underutilized Species ◽  
Recent Developments

Adapting to climate change, providing sufficient human food and nutritional needs, and securing sufficient energy supplies will call for a radical transformation from the current conventional adaptation approaches to more broad-based and transformative alternatives. This entails diversifying the agricultural system and boosting productivity of major cereal crops through development of climate-resilient cultivars that can sustainably maintain higher yields under climate change conditions, expanding our focus to crop wild relatives, and better exploitation of underutilized crop species. This is facilitated by the recent developments in plant genomics, such as advances in genome sequencing, assembly, and annotation, as well as gene editing technologies, which have increased the availability of high-quality reference genomes for various model and non-model plant species. This has necessitated genomics-assisted breeding of crops, including underutilized species, consequently broadening genetic variation of the available germplasm; improving the discovery of novel alleles controlling important agronomic traits; and enhancing creation of new crop cultivars with improved tolerance to biotic and abiotic stresses and superior nutritive quality. Here, therefore, we summarize these recent developments in plant genomics and their application, with particular reference to cereal crops (including underutilized species). Particularly, we discuss genome sequencing approaches, quantitative trait loci (QTL) mapping and genome-wide association (GWAS) studies, directed mutagenesis, plant non-coding RNAs, precise gene editing technologies such as CRISPR-Cas9, and complementation of crop genotyping by crop phenotyping. We then conclude by providing an outlook that, as we step into the future, high-throughput phenotyping, pan-genomics, transposable elements analysis, and machine learning hold much promise for crop improvements related to climate resilience and nutritional superiority.

scholarly journals Development of a Novel Orthotopic Gastric Cancer Mouse Model

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Wonyoung Kang ◽  
Leigh Maher ◽  
Michael Michaud ◽  
Seong-Woo Bae ◽  
Seongyeong Kim ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Mouse Models ◽  
Cancer Metastasis ◽  
Soft Tissues ◽  
Tumor Formation ◽  
Cancer Model ◽  
Orthotopic Model ◽  
Cancer Models ◽  
Organotropic Metastasis ◽  
Nsg Mouse

Abstract Background Gastric cancer metastasis is a highly fatal disease with a five-year survival rate of less than 5%. One major obstacle in studying gastric cancer metastasis is the lack of faithful models available. The cancer xenograft mouse models are widely used to elucidate the mechanisms of cancer development and progression. Current procedures for creating cancer xenografts include both heterotopic (i.e., subcutaneous) and orthotopic transplantation methods. Compared to the heterotopic model, the orthotopic model has been shown to be the more clinically relevant design as it enables the development of cancer metastasis. Although there are several methods in use to develop the orthotopic gastric cancer model, there is not a model which uses various types of tumor materials, such as soft tissues, semi-liquid tissues, or culture derivatives, due to the technical challenges. Thus, developing the applicable orthotopic model which can utilize various tumor materials is essential. Results To overcome the known limitations of the current orthotopic gastric cancer models, such as exposure of tumor fragments to the neighboring organs or only using firm tissues for the orthotopic implantation, we have developed a new method allowing for the complete insertion of soft tissue fragments or homogeneously minced tissues into the stomach submucosa layer of the immunodeficient NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mouse. With this completely-closed transplantation method, tumors with various types of tissue may be used to establish orthotopic gastric cancer models without the risks of exposure to nearby organs or cell leakage. This surgical procedure was highly reproducible in generating forty-eight mouse models with a surgery success rate of 96% and tumor formation of 93%. Among four orthotopic patient-derived xenograft (PDX) models that we generated in this study, we verified that the occurrence of organotropic metastasis in either the liver or peritoneal cavity was the same as that of the donor patients. Conclusion Here we describe a new protocol, step by step, for the establishment of orthotopic xenograft of gastric cancer. This novel technique will be able to increase the use of orthotopic models in broader applications for not only gastric cancer research but also any research related to the stomach microenvironment.

scholarly journals Mouse breast cancer model-dependent changes in metabolic syndrome-associated phenotypes caused by maternal dioxin exposure and dietary fat

2009 ◽  
Vol 296 (1) ◽  
pp. E203-E210 ◽  
Author(s):  
Michele La Merrill ◽  
David S. Baston ◽  
Michael S. Denison ◽  
Linda S. Birnbaum ◽  
Daniel Pomp ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Metabolic Syndrome ◽  
Adipose Tissue ◽  
Blood Glucose ◽  
Estrogenic Activity ◽  
Fasting Blood Glucose ◽  
Growth Spurt ◽  
Cancer Model ◽  
Serum Triglycerides ◽  
Cancer Models

Diets high in fat are associated with increased susceptibility to obesity and metabolic syndrome. Increased adipose tissue that is caused by high-fat diets (HFD) results in altered storage of lipophilic toxicants like 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD), which may further increase susceptibility to metabolic syndrome. Because both TCDD and HFD are associated with increased breast cancer risk, we examined their effects on metabolic syndrome-associated phenotypes in three mouse models of breast cancer: 7,12-dimethylbenz[a]anthracene (DMBA), Tg(MMTV-Neu)202Mul/J (HER2), and TgN(MMTV-PyMT)634Mul/J (PyMT), all on an FVB/N genetic background. Pregnant mice dosed with 1 μg/kg of TCDD or vehicle on gestational day 12.5 were placed on a HFD or low-fat diet (LFD) at parturition. Body weights, percent body fat, and fasting blood glucose were measured longitudinally, and triglycerides were measured at study termination. On HFD, all cancer models reached the pubertal growth spurt ahead of FVB controls. Among mice fed HFD, the HER2 model had a greater increase in body weight and adipose tissue from puberty through adulthood compared with the PyMT and DMBA models. However, the DMBA model consistently had higher fasting blood glucose levels than the PyMT and HER2 models. TCDD only impacted serum triglycerides in the PyMT model maintained on HFD. Because the estrogenic activity of the HFD was three times lower than that of the LFD, differential dietary estrogenic activities did not drive the observed phenotypic differences. Rather, the HFD-dependent changes were cancer model dependent. These results show that cancer models can have differential effects on metabolic syndrome-associated phenotypes even before cancers arise.

scholarly journals A Novel Assay for Profiling GBM Cancer Model Heterogeneity and Drug Screening

Cells ◽  
2019 ◽  
Vol 8 (7) ◽  
pp. 702 ◽  
Author(s):  
Christian T. Stackhouse ◽  
James R. Rowland ◽  
Rachael S. Shevin ◽  
Raj Singh ◽  
G. Yancey Gillespie ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Therapeutic Response ◽  
Specific Gene ◽  
Cancer Model ◽  
In Vivo Models ◽  
Cancer Models ◽  
Model Independent ◽  
Orthotopic Xenografts

Accurate patient-derived models of cancer are needed for profiling the disease and for testing therapeutics. These models must not only be accurate, but also suitable for high-throughput screening and analysis. Here we compare two derivative cancer models, microtumors and spheroids, to the gold standard model of patient-derived orthotopic xenografts (PDX) in glioblastoma multiforme (GBM). To compare these models, we constructed a custom NanoString panel of 350 genes relevant to GBM biology. This custom assay includes 16 GBM-specific gene signatures including a novel GBM subtyping signature. We profiled 11 GBM-PDX with matched orthotopic cells, derived microtumors, and derived spheroids using the custom NanoString assay. In parallel, these derivative models underwent drug sensitivity screening. We found that expression of certain genes were dependent on the cancer model while others were model-independent. These model-independent genes can be used in profiling tumor-specific biology and in gauging therapeutic response. It remains to be seen whether or not cancer model-specific genes may be directly or indirectly, through changes to tumor microenvironment, manipulated to improve the concordance of in vitro derivative models with in vivo models yielding better prediction of therapeutic response.

Co-operation of α-galactosylceramide-loaded tumour cells and TLR9 agonists induce potent anti-tumour responses in a murine colon cancer model

2015 ◽  
Vol 473 (1) ◽  
pp. 7-19 ◽  
Author(s):  
Tiangeng Dong ◽  
Tuo Yi ◽  
Mengxuan Yang ◽  
Shengli Lin ◽  
Wenxiang Li ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Tumour Cells ◽  
Toll Like Receptor ◽  
Cancer Model ◽  
Cancer Vaccination ◽  
Tlr9 Agonist ◽  
Broad Applicability ◽  
Human Cancers ◽  
Anti Cancer ◽  
Murine Colon

This is the first demonstration that α-galactosylceramide (α-Galcer)-loaded tumour cells (tumour–Gal) plus the Toll-like receptor 9 (TLR9) agonist as therapeutic anti-cancer vaccination. This strategy may have broad applicability and lead to development of effective immunotherapy against various human cancers.

Microbiota Effects on Carcinogenesis: Initiation, Promotion and Progression

2020 ◽  
Vol 72 (1) ◽  
Author(s):  
Lacey R. Lopez ◽  
Rachel M. Bleich ◽  
Janelle C. Arthur
Keyword(s):  
Immune Evasion ◽  
Fusobacterium Nucleatum ◽  
Cancer Development ◽  
Annual Review ◽  
Publication Date ◽  
Epigenetic Changes ◽  
Microbial Activities ◽  
Normal Cells ◽  
Multistep Process ◽  
Host Genetic

Carcinogenesis is a multistep process by which normal cells acquire genetic and epigenetic changes that result in cancer. In combination with host genetic susceptibility and environmental exposures, a prominent procarcinogenic role for the microbiota has recently emerged. In colorectal cancer (CRC), three nefarious microbes have been consistently linked to cancer development: ( a) Colibactin-producing Escherichia coli initiates carcinogenic DNA damage, ( b) enterotoxigenic Bacteroides fragilis promotes tumorigenesis via toxin-induced cell proliferation and tumor-promoting inflammation, and ( c) Fusobacterium nucleatum enhances CRC progression through two adhesins, Fap2 and FadA, that promote proliferation and antitumor immune evasion and may contribute to metastases. Herein, we use these three prominent microbes to discuss the experimental evidence linking microbial activities to carcinogenesis and the specific mechanisms driving this stepwise process. Precisely defining mechanisms by which the microbiota impacts carcinogenesis at each stage is essential for developing microbiota-targeted strategies for the diagnosis, prognosis, and treatment of cancer. Expected final online publication date for the Annual Review of Medicine, Volume 72 is January 27, 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

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