Frontiers in genome editing最新文献_第5页

Hs1Cas12a and Ev1Cas12a confer efficient genome editing in plants. Hs1Cas12a和Ev1Cas12a赋予植物有效的基因组编辑。

Frontiers in genome editing Pub Date : 2023-10-12 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1251903

Gen Li, Yingxiao Zhang, Micah Dailey, Yiping Qi

引用次数: 0

How genome editing changed the world of large animal research. 基因组编辑如何改变了大型动物研究的世界。

Frontiers in genome editing Pub Date : 2023-10-11 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1272687

Konrad Fischer, Angelika Schnieke

{"title":"How genome editing changed the world of large animal research.","authors":"Konrad Fischer, Angelika Schnieke","doi":"10.3389/fgeed.2023.1272687","DOIUrl":"10.3389/fgeed.2023.1272687","url":null,"abstract":"The first genetically modified large animals were developed in 1985 by microinjection to increase the growth of agricultural livestock such as pigs. Since then, it has been a difficult trail due to the lack of genetic tools. Although methods and technologies were developed quickly for the main experimental mammal, the mouse, e.g., efficient pronuclear microinjection, gene targeting in embryonic stem cells, and omics data, most of it was-and in part still is-lacking when it comes to livestock. Over the next few decades, progress in genetic engineering of large animals was driven less by research for agriculture but more for biomedical applications, such as the production of pharmaceutical proteins in the milk of sheep, goats, or cows, xeno-organ transplantation, and modeling human diseases. Available technologies determined if a desired animal model could be realized, and efficiencies were generally low. Presented here is a short review of how genome editing tools, specifically CRISPR/Cas, have impacted the large animal field in recent years. Although there will be a focus on genome engineering of pigs for biomedical applications, the general principles and experimental approaches also apply to other livestock species or applications.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"5 ","pages":"1272687"},"PeriodicalIF":0.0,"publicationDate":"2023-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10598601/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54232727","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

Recalcitrance to transformation, a hindrance for genome editing of legumes. 对转化的抗拒，是豆类基因组编辑的障碍。

IF 4.9

Frontiers in genome editing Pub Date : 2023-09-21 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1247815

V M Nivya, Jasmine M Shah

{"title":"Recalcitrance to transformation, a hindrance for genome editing of legumes.","authors":"V M Nivya, Jasmine M Shah","doi":"10.3389/fgeed.2023.1247815","DOIUrl":"10.3389/fgeed.2023.1247815","url":null,"abstract":"Plant genome editing, a recently discovered method for targeted mutagenesis, has emerged as a promising tool for crop improvement and gene function research. Many genome-edited plants, such as rice, wheat, and tomato, have emerged over the last decade. As the preliminary steps in the procedure for genome editing involve genetic transformation, amenability to genome editing depends on the efficiency of genetic engineering. Hence, there are numerous reports on the aforementioned crops because they are transformed with relative ease. Legume crops are rich in protein and, thus, are a favored source of plant proteins for the human diet in most countries. However, legume cultivation often succumbs to various biotic/abiotic threats, thereby leading to high yield loss. Furthermore, certain legumes like peanuts possess allergens, and these need to be eliminated as these deprive many people from gaining the benefits of such crops. Further genetic variations are limited in certain legumes. Genome editing has the potential to offer solutions to not only combat biotic/abiotic stress but also generate desirable knock-outs and genetic variants. However, excluding soybean, alfalfa, and Lotus japonicus, reports obtained on genome editing of other legume crops are less. This is because, excluding the aforementioned three legume crops, the transformation efficiency of most legumes is found to be very low. Obtaining a higher number of genome-edited events is desirable as it offers the option to genotypically/phenotypically select the best candidate, without the baggage of off-target mutations. Eliminating the barriers to genetic engineering would directly help in increasing genome-editing rates. Thus, this review aims to compare various legumes for their transformation, editing, and regeneration efficiencies and discusses various solutions available for increasing transformation and genome-editing rates in legumes.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"5 ","pages":"1247815"},"PeriodicalIF":4.9,"publicationDate":"2023-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10551638/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41175205","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-wide CRISPR screens and their applications in infectious disease. 全基因组CRISPR筛选及其在传染病中的应用。

Frontiers in genome editing Pub Date : 2023-09-19 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1243731

Kaveri Srivastava, Bhaswati Pandit

{"title":"Genome-wide CRISPR screens and their applications in infectious disease.","authors":"Kaveri Srivastava, Bhaswati Pandit","doi":"10.3389/fgeed.2023.1243731","DOIUrl":"https://doi.org/10.3389/fgeed.2023.1243731","url":null,"abstract":"Inactivation or targeted disruption of a gene provides clues to assess the function of the gene in many cellular processes. Knockdown or knocking out a gene has been widely used for this purpose. However, recently CRISPR mediated genome editing has taken over the knockout/knockdown system with more precision. CRISPR technique has enabled us to perform targeted mutagenesis or genome editing to address questions in fundamental biology to biomedical research. Its application is wide in understanding the role of genes in the disease process, and response to therapy in cancer, metabolic disorders, or infectious disease. In this article, we have focused on infectious disease and how genome-wide CRISPR screens have enabled us to identify host factors involved in the process of infection. Understanding the biology of the host-pathogen interaction is of immense importance in planning host-directed therapy to improve better management of the disease. Genome-wide CRISPR screens provide strong mechanistic ways to identify the host dependency factors involved in various infections. We presented insights into genome-wide CRISPR screens conducted in the context of infectious diseases both viral and bacterial that led to better understanding of host-pathogen interactions and immune networks. We have discussed the advancement of knowledge pertaining to influenza virus, different hepatitis viruses, HIV, most recent SARS CoV2 and few more. Among bacterial diseases, we have focused on infection with life threatening Mycobacteria, Salmonella, S. aureus, etc. It appears that the CRISPR technique can be applied universally to multiple infectious disease models to unravel the role of known or novel host factors.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"5 ","pages":"1243731"},"PeriodicalIF":0.0,"publicationDate":"2023-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10546192/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41164720","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

Editorial: Genome edited organisms for agriculture-challenges and perspectives for development and regulation. 社论：基因组编辑的生物对农业的挑战和发展和监管的前景。

Frontiers in genome editing Pub Date : 2023-09-18 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1287973

Michael Eckerstorfer, Sarah Zanon Agapito-Tenfen, Gijs A Kleter

引用次数: 0

Current approaches and potential challenges in the delivery of gene editing cargos into hematopoietic stem and progenitor cells. 将基因编辑货物递送到造血干细胞和祖细胞中的当前方法和潜在挑战。

Frontiers in genome editing Pub Date : 2023-09-15 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1148693

Ramya Murugesan, Karthik V Karuppusamy, Srujan Marepally, Saravanabhavan Thangavel

引用次数: 0

Efficient DNA knock-in using AAV-mediated delivery with 2-cell embryo CRISPR-Cas9 electroporation. 利用 AAV 介导的传递与 2 细胞胚胎 CRISPR-Cas9 电穿孔技术实现高效 DNA 基因敲入。

Frontiers in genome editing Pub Date : 2023-08-25 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1256451

Daniel J Davis, James F McNew, Hailey Maresca-Fichter, Kaiwen Chen, Bhanu P Telugu, Elizabeth C Bryda

{"title":"Efficient DNA knock-in using AAV-mediated delivery with 2-cell embryo CRISPR-Cas9 electroporation.","authors":"Daniel J Davis, James F McNew, Hailey Maresca-Fichter, Kaiwen Chen, Bhanu P Telugu, Elizabeth C Bryda","doi":"10.3389/fgeed.2023.1256451","DOIUrl":"10.3389/fgeed.2023.1256451","url":null,"abstract":"Recent advances in CRISPR-Cas genome editing technology have been instrumental in improving the efficiency to produce genetically modified animal models. In this study we have combined four very promising approaches to come up with a highly effective pipeline to produce knock-in mouse and rat models. The four combined methods include: AAV-mediated DNA delivery, single-stranded DNA donor templates, 2-cell embryo modification, and CRISPR-Cas ribonucleoprotein (RNP) electroporation. Using this new combined approach, we were able to produce successfully targeted knock-in rat models containing either Cre or Flp recombinase sequences with knock-in efficiencies over 90%. Furthermore, we were able to produce a knock-in mouse model containing a Cre recombinase targeted insertion with over 50% knock-in efficiency directly comparing efficiencies to other commonly used approaches. Our modified AAV-mediated DNA delivery with 2-cell embryo CRISPR-Cas9 RNP electroporation technique has proven to be highly effective for generating both knock-in mouse and knock-in rat models.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"5 ","pages":"1256451"},"PeriodicalIF":0.0,"publicationDate":"2023-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10485772/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10220069","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

Potentials of genotypes, morpho-physio-biochemical traits, and growing media on shelf life and future prospects of gene editing in tomatoes. 基因型、形态生理生化性状和生长介质对番茄货架期的影响及基因编辑的未来前景。

IF 4.9

Frontiers in genome editing Pub Date : 2023-08-23 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1203485

Renu Yadav, Sarika Jaiswal, Tripti Singhal, Rohit Kumar Mahto, S B Verma, Ramesh Kumar Yadav, Rajendra Kumar

{"title":"Potentials of genotypes, morpho-physio-biochemical traits, and growing media on shelf life and future prospects of gene editing in tomatoes.","authors":"Renu Yadav, Sarika Jaiswal, Tripti Singhal, Rohit Kumar Mahto, S B Verma, Ramesh Kumar Yadav, Rajendra Kumar","doi":"10.3389/fgeed.2023.1203485","DOIUrl":"10.3389/fgeed.2023.1203485","url":null,"abstract":"Background: To study the genetic basis of the impact of genotypes and morpho-physio-biochemical traits under different organic and inorganic fertilizer doses on the shelf life attribute of tomatoes, field experiments were conducted in randomized block designs during the rabi seasons of 2018-2019 and 2019-2020. The experiment comprised three diverse nutrient environments [T1-organic; T2-inorganic; T3-control (without any fertilizers)] and five tomato genotypes with variable growth habits, specifically Angoorlata (Indeterminate), Avinash-3 (semi-determinate), Swaraksha (semi-determinate), Pusa Sheetal (semi-determinate), and Pusa Rohini (determinate). Results: The different tomato genotypes behaved apparently differently from each other in terms of shelf life. All the genotypes had maximum shelf life when grown in organic environments. However, the Pusa Sheetal had a maximum shelf life of 8.35 days when grown in an organic environment and showed an increase of 12% over the control. The genotype Pusa Sheetal, organic environment and biochemical trait Anthocyanin provides a promise as potential contributor to improve the keeping quality of tomatoes. Conclusion: The genotype Pusa Sheetal a novel source for shelf life, organic environment, and anthocyanin have shown promises for extended shelf life in tomatoes. Thus, the identified trait and genotype can be utilized in tomato improvement programs. Furthermore, this identified trait can also be targeted for its quantitative enhancement in order to increase tomato shelf life through a genome editing approach. A generalized genome editing mechanism is consequently suggested.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"5 ","pages":"1203485"},"PeriodicalIF":4.9,"publicationDate":"2023-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10481343/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10184973","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

Genetic manipulation of betta fish. betta 鱼的遗传操作。

Frontiers in genome editing Pub Date : 2023-07-21 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1167093

Alec Palmiotti, Madison R Lichak, Pei-Yin Shih, Young Mi Kwon, Andres Bendesky

{"title":"Genetic manipulation of betta fish.","authors":"Alec Palmiotti, Madison R Lichak, Pei-Yin Shih, Young Mi Kwon, Andres Bendesky","doi":"10.3389/fgeed.2023.1167093","DOIUrl":"10.3389/fgeed.2023.1167093","url":null,"abstract":"Betta splendens, also known as Siamese fighting fish or \"betta,\" is a freshwater fish species renowned for its astonishing morphological diversity and extreme aggressive behavior. Despite recent advances in our understanding of the genetics and neurobiology of betta, the lack of tools to manipulate their genome has hindered progress at functional and mechanistic levels. In this study, we outline the use of three genetic manipulation technologies, which we have optimized for use in betta: CRISPR/Cas9-mediated knockout, CRISPR/Cas9-mediated knockin, and Tol2-mediated transgenesis. We knocked out three genes: alkal2l, bco1l, and mitfa, and analyzed their effects on viability and pigmentation. Furthermore, we knocked in a fluorescent protein into the mitfa locus, a proof-of-principle experiment of this powerful technology in betta. Finally, we used Tol2-mediated transgenesis to create fish with ubiquitous expression of GFP, and then developed a bicistronic plasmid with heart-specific expression of a red fluorescent protein to serve as a visible marker of successful transgenesis. Our work highlights the potential for the genetic manipulation of betta, providing valuable resources for the effective use of genetic tools in this animal model.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"5 ","pages":"1167093"},"PeriodicalIF":0.0,"publicationDate":"2023-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10401044/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10325328","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

Strategies for delivery of CRISPR/Cas-mediated genome editing to obtain edited plants directly without transgene integration. 提供 CRISPR/Cas 介导的基因组编辑的策略，以直接获得编辑过的植物，而无需转基因整合。

IF 4.9

Frontiers in genome editing Pub Date : 2023-07-20 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1209586

Zuzana Kocsisova, Viktoriya Coneva

{"title":"Strategies for delivery of CRISPR/Cas-mediated genome editing to obtain edited plants directly without transgene integration.","authors":"Zuzana Kocsisova, Viktoriya Coneva","doi":"10.3389/fgeed.2023.1209586","DOIUrl":"10.3389/fgeed.2023.1209586","url":null,"abstract":"Increased understanding of plant genetics and the development of powerful and easier-to-use gene editing tools over the past century have revolutionized humankind's ability to deliver precise genotypes in crops. Plant transformation techniques are well developed for making transgenic varieties in certain crops and model organisms, yet reagent delivery and plant regeneration remain key bottlenecks to applying the technology of gene editing to most crops. Typical plant transformation protocols to produce transgenic, genetically modified (GM) varieties rely on transgenes, chemical selection, and tissue culture. Typical protocols to make gene edited (GE) varieties also use transgenes, even though these may be undesirable in the final crop product. In some crops, the transgenes are routinely segregated away during meiosis by performing crosses, and thus only a minor concern. In other crops, particularly those propagated vegetatively, complex hybrids, or crops with long generation times, such crosses are impractical or impossible. This review highlights diverse strategies to deliver CRISPR/Cas gene editing reagents to regenerable plant cells and to recover edited plants without unwanted integration of transgenes. Some examples include delivering DNA-free gene editing reagents such as ribonucleoproteins or mRNA, relying on reagent expression from non-integrated DNA, using novel delivery mechanisms such as viruses or nanoparticles, using unconventional selection methods to avoid integration of transgenes, and/or avoiding tissue culture altogether. These methods are advancing rapidly and already enabling crop scientists to make use of the precision of CRISPR gene editing tools.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"5 ","pages":"1209586"},"PeriodicalIF":4.9,"publicationDate":"2023-07-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10398581/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10005816","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