Frontiers in genome editing最新文献_第9页

Canadian Consumer Preferences Regarding Gene-Edited Food Products 加拿大消费者对基因编辑食品的偏好

Frontiers in genome editing Pub Date : 2022-04-11 DOI: 10.3389/fgeed.2022.854334

Oswaldo Vasquez, Hayley Hesseln, S. Smyth

{"title":"Canadian Consumer Preferences Regarding Gene-Edited Food Products","authors":"Oswaldo Vasquez, Hayley Hesseln, S. Smyth","doi":"10.3389/fgeed.2022.854334","DOIUrl":"https://doi.org/10.3389/fgeed.2022.854334","url":null,"abstract":"Innovations in food production and processing have largely remained “behind the scenes” for decades. The current nature of social media and calls for increased transparency regarding food results in a new landscape where consumer product demands are more important than ever, but are increasingly based on limited, or incorrect, information. One area where consumer awareness is rapidly emerging is the area of gene-edited food products. This article uses a consumer survey to gather perceptions regarding food safety, gene editing and willingness to consume for three gene-edited food products. Four factors were found to strongly influence consumer perceptions: trust in the Canadian food safety system; food technology neophobia scores; knowledge of genetics; and self-knowledge of gene editing. The survey of 497 Canadians found that 15% identified as neophobics and 12% as neophilics. The majority of participants identified as neutral. When presented with various food values, participants indicated that nutrition, price, and taste were the three most important values. A participants’ willingness to consume gene-edited food products strongly correlated with neophobic and neophilic preferences, with neophobics unwilling to consume and neophilics being uncertain. The only food value that strongly affects consumer willingness to consume is the environmental impact of a products’ production. Canadian consumers have a moderate to high level of trust in Canada’s food safety system, but this level of trust fails to carry over to food products produced through innovative technologies; however, consumers express a higher level of trust in gene-edited technology than genetically modified technology.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41852975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Imaging Unique DNA Sequences in Individual Cells Using a CRISPR-Cas9-Based, Split Luciferase Biosensor 使用基于CRISPR-Cas9的分裂萤光素酶生物传感器成像单个细胞中的独特DNA序列

Frontiers in genome editing Pub Date : 2022-03-25 DOI: 10.3389/fgeed.2022.867390

Nicholas G. Heath, H. O’Geen, Nicole B. Halmai, J. Corn, D. Segal

{"title":"Imaging Unique DNA Sequences in Individual Cells Using a CRISPR-Cas9-Based, Split Luciferase Biosensor","authors":"Nicholas G. Heath, H. O’Geen, Nicole B. Halmai, J. Corn, D. Segal","doi":"10.3389/fgeed.2022.867390","DOIUrl":"https://doi.org/10.3389/fgeed.2022.867390","url":null,"abstract":"An extensive arsenal of biosensing tools has been developed based on the clustered regularly interspaced short palindromic repeat (CRISPR) platform, including those that detect specific DNA sequences both in vitro and in live cells. To date, DNA imaging approaches have traditionally used full fluorescent reporter-based fusion probes. Such “always-on” probes differentiate poorly between bound and unbound probe and are unable to sensitively detect unique copies of a target sequence in individual cells. Herein we describe a DNA biosensor that provides a sensitive readout for such low-copy DNA sequences through proximity-mediated reassembly of two independently optimized fragments of NanoLuc luciferase (NLuc), a small, bright luminescent reporter. Applying this “turn-on” probe in live cells, we demonstrate an application not easily achieved by fluorescent reporter-based probes, detection of individual endogenous genomic loci using standard epifluorescence microscopy. This approach could enable detection of gene edits during ex vivo editing procedures and should be a useful platform for many other live cell DNA biosensing applications.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48320541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Rapid Assessment of CRISPR Transfection Efficiency and Enrichment of CRISPR Induced Mutations Using a Dual-Fluorescent Stable Reporter System 使用双荧光稳定报告系统快速评估CRISPR转染效率和富集CRISPR诱导的突变

Frontiers in genome editing Pub Date : 2022-03-21 DOI: 10.3389/fgeed.2022.854866

Karim E Shalaby, Mustapha Aouida, Vijay Gupta, S. Ghanem, O. El‐Agnaf

{"title":"Rapid Assessment of CRISPR Transfection Efficiency and Enrichment of CRISPR Induced Mutations Using a Dual-Fluorescent Stable Reporter System","authors":"Karim E Shalaby, Mustapha Aouida, Vijay Gupta, S. Ghanem, O. El‐Agnaf","doi":"10.3389/fgeed.2022.854866","DOIUrl":"https://doi.org/10.3389/fgeed.2022.854866","url":null,"abstract":"The nuclease activity of the CRISPR-Cas9 system relies on the delivery of a CRISPR-associated protein 9 (Cas9) and a single guide RNA (sgRNA) against the target gene. CRISPR components are typically delivered to cells as either a Cas9/sgRNA ribonucleoprotein (RNP) complex or a plasmid encoding a Cas9 protein along with a sequence-specific sgRNA. Multiple transfection reagents are known to deliver CRISPR-Cas9 components, and delivery vectors are being developed for different purposes by several groups. Here, we repurposed a dual-fluorescence (RFP-GFP-GFP) reporter system to quantify the uptake level of the functional CRISPR-Cas9 components into cells and compare the efficiency of CRISPR delivery vectors. Using this system, we developed a novel and rapid cell-based microplate reader assay that makes possible real-time, rapid, and high throughput quantification of CRISPR nuclease activity. Cells stably expressing this dual-fluorescent reporter construct facilitated a direct quantification of the level of the internalized and functional CRISPR-Cas9 molecules into the cells without the need of co-transfecting fluorescently labeled reporter molecules. Additionally, targeting a reporter gene integrated into the genome recapitulates endogenous gene targeting. Thus, this reporter could be used to optimize various transfection conditions of CRISPR components, to evaluate and compare the efficiency of transfection agents, and to enrich cells containing desired CRISPR-induced mutations.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47822044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Contributions of Genome Editing Technologies Towards Improved Nutrition, Environmental Sustainability and Poverty Reduction. 基因组编辑技术对改善营养、环境可持续性和减少贫困的贡献。

IF 4.9

Frontiers in genome editing Pub Date : 2022-03-17 eCollection Date: 2022-01-01 DOI: 10.3389/fgeed.2022.863193

Stuart J Smyth

{"title":"Contributions of Genome Editing Technologies Towards Improved Nutrition, Environmental Sustainability and Poverty Reduction.","authors":"Stuart J Smyth","doi":"10.3389/fgeed.2022.863193","DOIUrl":"10.3389/fgeed.2022.863193","url":null,"abstract":"The Sustainable Development Goals (SDGs) were launched in 2015, with the top three goals being poverty eradication, improved food security and increased human health. All 17 SDGs have a target achievement date of 2030. These are ambitious and inspirational goals that require substantial innovation and technology adoption for successful achievement. Innovations in plant breeding have substantially contributed to transforming the efficiency of food production since the mid 20th century, with innovations emerging in the current millennium demonstrating enhanced potential to improve crop yields, the nutritional values of food crops and environmental impacts. These outcomes underpin several SDGs, but in particular the first three. As climate change is expected to become increasingly variable, with greater impacts on agriculture, the ability to ensure increased food production is going to be increasingly important, as higher yields directly contribute to reducing poverty. This article reviews recent reports of potential contributions from genome editing technologies in terms of increased yield, enhanced nutrition and greater sustainability, highlighting their importance for achieving the leading three SDGs.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 1","pages":"863193"},"PeriodicalIF":4.9,"publicationDate":"2022-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8968197/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69928227","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

CRISPR/Cas-Based Gene Editing Strategies for DOCK8 Immunodeficiency Syndrome. 基于CRISPR/ cas的DOCK8免疫缺陷综合征基因编辑策略

IF 4.9

Frontiers in genome editing Pub Date : 2022-03-17 eCollection Date: 2022-01-01 DOI: 10.3389/fgeed.2022.793010

Sujan Ravendran, Sabina Sánchez Hernández, Saskia König, Rasmus O Bak

{"title":"CRISPR/Cas-Based Gene Editing Strategies for DOCK8 Immunodeficiency Syndrome.","authors":"Sujan Ravendran, Sabina Sánchez Hernández, Saskia König, Rasmus O Bak","doi":"10.3389/fgeed.2022.793010","DOIUrl":"10.3389/fgeed.2022.793010","url":null,"abstract":"Defects in the DOCK8 gene causes combined immunodeficiency termed DOCK8 immunodeficiency syndrome (DIDS). DIDS previously belonged to the disease category of autosomal recessive hyper IgE syndrome (AR-HIES) but is now classified as a combined immunodeficiency (CID). This genetic disorder induces early onset of susceptibility to severe recurrent viral and bacterial infections, atopic diseases and malignancy resulting in high morbidity and mortality. This pathological state arises from impairment of actin polymerization and cytoskeletal rearrangement, which induces improper immune cell migration-, survival-, and effector functions. Owing to the severity of the disease, early allogenic hematopoietic stem cell transplantation is recommended even though it is associated with risk of unintended adverse effects, the need for compatible donors, and high expenses. So far, no alternative therapies have been developed, but the monogenic recessive nature of the disease suggests that gene therapy may be applied. The advent of the CRISPR/Cas gene editing system heralds a new era of possibilities in precision gene therapy, and positive results from clinical trials have already suggested that the tool may provide definitive cures for several genetic disorders. Here, we discuss the potential application of different CRISPR/Cas-mediated genetic therapies to correct the DOCK8 gene. Our findings encourage the pursuit of CRISPR/Cas-based gene editing approaches, which may constitute more precise, affordable, and low-risk definitive treatment options for DOCK8 deficiency.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 1","pages":"793010"},"PeriodicalIF":4.9,"publicationDate":"2022-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8969908/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41480667","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Base Editors for Citrus Gene Editing. 柑橘基因编辑的碱基编辑器

IF 4.9

Frontiers in genome editing Pub Date : 2022-02-28 eCollection Date: 2022-01-01 DOI: 10.3389/fgeed.2022.852867

Xiaoen Huang, Yuanchun Wang, Nian Wang

{"title":"Base Editors for Citrus Gene Editing.","authors":"Xiaoen Huang, Yuanchun Wang, Nian Wang","doi":"10.3389/fgeed.2022.852867","DOIUrl":"10.3389/fgeed.2022.852867","url":null,"abstract":"Base editors, such as adenine base editors (ABE) and cytosine base editors (CBE), provide alternatives for precise genome editing without generating double-strand breaks (DSBs), thus avoiding the risk of genome instability and unpredictable outcomes caused by DNA repair. Precise gene editing mediated by base editors in citrus has not been reported. Here, we have successfully adapted the ABE to edit the TATA box in the promoter region of the canker susceptibility gene LOB1 from TATA to CACA in grapefruit (Citrus paradise) and sweet orange (Citrus sinensis). TATA-edited plants are resistant to the canker pathogen Xanthomonas citri subsp. citri (Xcc). In addition, CBE was successfully used to edit the acetolactate synthase (ALS) gene in citrus. ALS-edited plants were resistant to the herbicide chlorsulfuron. Two ALS-edited plants did not show green fluorescence although the starting construct for transformation contains a GFP expression cassette. The Cas9 gene was undetectable in the herbicide-resistant citrus plants. This indicates that the ALS edited plants are transgene-free, representing the first transgene-free gene-edited citrus using the CRISPR technology. In summary, we have successfully adapted the base editors for precise citrus gene editing. The CBE base editor has been used to generate transgene-free citrus via transient expression.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 1","pages":"852867"},"PeriodicalIF":4.9,"publicationDate":"2022-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8919994/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41630584","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Genome Editing for Improving Crop Nutrition 基因组编辑改善作物营养

Frontiers in genome editing Pub Date : 2022-02-09 DOI: 10.3389/fgeed.2022.850104

Ai Nagamine, H. Ezura

{"title":"Genome Editing for Improving Crop Nutrition","authors":"Ai Nagamine, H. Ezura","doi":"10.3389/fgeed.2022.850104","DOIUrl":"https://doi.org/10.3389/fgeed.2022.850104","url":null,"abstract":"Genome editing technologies, including CRISPR/Cas9 and TALEN, are excellent genetic modification techniques and are being proven to be powerful tools not only in the field of basic science but also in the field of crop breeding. Recently, two genome-edited crops targeted for nutritional improvement, high GABA tomatoes and high oleic acid soybeans, have been released to the market. Nutritional improvement in cultivated crops has been a major target of conventional genetic modification technologies as well as classical breeding methods. Mutations created by genome editing are considered to be almost identical to spontaneous genetic mutations because the mutation inducer, the transformed foreign gene, can be completely eliminated from the final genome-edited hosts after causing the mutation. Therefore, genome-edited crops are expected to be relatively easy to supply to the market, unlike GMO crops. On the other hand, due to their technical feature, the main goal of current genome-edited crop creation is often the total or partial disruption of genes rather than gene delivery. Therefore, to obtain the desired trait using genome editing technology, in some cases, a different approach from that of genetic recombination technology may be required. In this mini-review, we will review several nutritional traits in crops that have been considered suitable targets for genome editing, including the two examples mentioned above, and discuss how genome editing technology can be an effective breeding technology for improving nutritional traits in crops.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46776909","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

Genome editing approaches using reproductive cells/tissues in flowering plants. 利用开花植物生殖细胞/组织的基因组编辑方法。

Frontiers in genome editing Pub Date : 2022-01-01 DOI: 10.3389/fgeed.2022.1085023

Erika Toda, Norio Kato, Tetsuya Higashiyama, Takashi Okamoto

{"title":"Genome editing approaches using reproductive cells/tissues in flowering plants.","authors":"Erika Toda, Norio Kato, Tetsuya Higashiyama, Takashi Okamoto","doi":"10.3389/fgeed.2022.1085023","DOIUrl":"https://doi.org/10.3389/fgeed.2022.1085023","url":null,"abstract":"Targeted mutagenesis via programmable nucleases including the clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) (CRISPR/Cas9) system has been broadly utilized to generate genome-edited organisms including flowering plants. To date, specific expression of Cas9 protein and guide RNA (gRNA) in reproductive cells or tissues is considered one of the most effective genome-editing approaches for heritable targeted mutagenesis. In this report, we review recent advances in genome editing methods for reproductive cells or tissues, which have roles in transmitting genetic material to the next-generation, such as egg cells, pollen grains, zygotes, immature zygotic embryos, and shoot apical meristems (SAMs). Specific expression of Cas9 proteins in initiating cells efficiently induces targeted mutagenesis via Agrobacterium-mediated in planta transformation. In addition, genome editing by direct delivery of CRISPR/Cas9 components into pollen grains, zygotes, cells of embryos and SAMs has been successfully established to generate genome-edited plant lines. Notably, DNA-free genome editing by the delivery of Cas9-gRNA ribonucleoproteins (RNPs) is not associated with any legislative concerns about genetically modified organisms. In summary, the genome editing methods for reproductive cells or tissues have enormous potential for not only basic studies for plant reproduction but also applied sciences toward molecular plant breeding.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"1085023"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9873966/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10584240","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Corrigendum: CRISPR nuclease off-target activity and mitigation strategies. 勘误:CRISPR核酸酶脱靶活性和缓解策略。

Frontiers in genome editing Pub Date : 2022-01-01 DOI: 10.3389/fgeed.2022.1112956

Beeke Wienert, M Kyle Cromer

引用次数: 1

Hematopoietic stem and progenitors cells gene editing: Beyond blood disorders. 造血干细胞和祖细胞基因编辑:超越血液病。

Frontiers in genome editing Pub Date : 2022-01-01 DOI: 10.3389/fgeed.2022.997142

Valentina Buffa, José Roberto Alvarez Vargas, Anne Galy, Simone Spinozzi, Céline J Rocca

{"title":"Hematopoietic stem and progenitors cells gene editing: Beyond blood disorders.","authors":"Valentina Buffa, José Roberto Alvarez Vargas, Anne Galy, Simone Spinozzi, Céline J Rocca","doi":"10.3389/fgeed.2022.997142","DOIUrl":"https://doi.org/10.3389/fgeed.2022.997142","url":null,"abstract":"Lessons learned from decades-long practice in the transplantation of hematopoietic stem and progenitor cells (HSPCs) to treat severe inherited disorders or cancer, have set the stage for the current ex vivo gene therapies using autologous gene-modified hematopoietic stem and progenitor cells that have treated so far, hundreds of patients with monogenic disorders. With increased knowledge of hematopoietic stem and progenitor cell biology, improved modalities for patient conditioning and with the emergence of new gene editing technologies, a new era of hematopoietic stem and progenitor cell-based gene therapies is poised to emerge. Gene editing has the potential to restore physiological expression of a mutated gene, or to insert a functional gene in a precise locus with reduced off-target activity and toxicity. Advances in patient conditioning has reduced treatment toxicities and may improve the engraftment of gene-modified cells and specific progeny. Thanks to these improvements, new potential treatments of various blood- or immune disorders as well as other inherited diseases will continue to emerge. In the present review, the most recent advances in hematopoietic stem and progenitor cell gene editing will be reported, with a focus on how this approach could be a promising solution to treat non-blood-related inherited disorders and the mechanisms behind the therapeutic actions discussed.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"997142"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9868335/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9177522","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2