Frontiers in genome editing最新文献_第5页

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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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

Methods of crop improvement and applications towards fortifying food security. 作物改良方法及其在加强粮食安全方面的应用。

Frontiers in genome editing Pub Date : 2023-07-07 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1171969

Aayushi Patel, Andrew Miles, Tara Strackhouse, Logan Cook, Sining Leng, Shrina Patel, Kelsey Klinger, Sairam Rudrabhatla, Shobha D Potlakayala

{"title":"Methods of crop improvement and applications towards fortifying food security.","authors":"Aayushi Patel, Andrew Miles, Tara Strackhouse, Logan Cook, Sining Leng, Shrina Patel, Kelsey Klinger, Sairam Rudrabhatla, Shobha D Potlakayala","doi":"10.3389/fgeed.2023.1171969","DOIUrl":"10.3389/fgeed.2023.1171969","url":null,"abstract":"Agriculture has supported human life from the beginning of civilization, despite a plethora of biotic (pests, pathogens) and abiotic (drought, cold) stressors being exerted on the global food demand. In the past 50 years, the enhanced understanding of cellular and molecular mechanisms in plants has led to novel innovations in biotechnology, resulting in the introduction of desired genes/traits through plant genetic engineering. Targeted genome editing technologies such as Zinc-Finger Nucleases (ZFNs), Transcription Activator-Like Effector Nucleases (TALENs), and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) have emerged as powerful tools for crop improvement. This new CRISPR technology is proving to be an efficient and straightforward process with low cost. It possesses applicability across most plant species, targets multiple genes, and is being used to engineer plant metabolic pathways to create resistance to pathogens and abiotic stressors. These novel genome editing (GE) technologies are poised to meet the UN's sustainable development goals of \"zero hunger\" and \"good human health and wellbeing.\" These technologies could be more efficient in developing transgenic crops and aid in speeding up the regulatory approvals and risk assessments conducted by the US Departments of Agriculture (USDA), Food and Drug Administration (FDA), and Environmental Protection Agency (EPA).","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10361821/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10241066","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

Exploring the future of GM technology in sustainable local food systems in Colombia. 探索转基因技术在哥伦比亚可持续地方粮食系统中的未来。

IF 4.9

Frontiers in genome editing Pub Date : 2023-06-30 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1181811

Néstor Julián Cárdenas Pardo, Dolly Esperanza Rodriguez Robayo, John Cristhian Fernandez Lizarazo, Diego Camilo Peña-Quemba, Erica McGale

{"title":"Exploring the future of GM technology in sustainable local food systems in Colombia.","authors":"Néstor Julián Cárdenas Pardo, Dolly Esperanza Rodriguez Robayo, John Cristhian Fernandez Lizarazo, Diego Camilo Peña-Quemba, Erica McGale","doi":"10.3389/fgeed.2023.1181811","DOIUrl":"10.3389/fgeed.2023.1181811","url":null,"abstract":"The security of Earth's food systems is challenged by shifting regional climates. While agricultural processes are disrupted by climate change, they also play a large role in contributing to destabilizing greenhouse gases. Finding new strategies to increase yields while decreasing agricultural environmental impacts is essential. Tropical agriculture is particularly susceptible to climate change: local, smallholder farming, which provides a majority of the food supply, is high risk and has limited adaptation capacity. Rapid, inexpensive, intuitive solutions are needed, like the implementation of genetically modified (GM) crops. In the Latin American tropics, high awareness and acceptance of GM technologies, opportunities to test GM crops as part of local agricultural educations, and their known economic benefits, support their use. However, this is not all that is needed for the future of GM technologies in these areas: GM implementation must also consider environmental and social sustainability, which can be unique to a locality. Primarily from the perspective of its educators, the potential of a rural Colombian university in driving GM implementation is explored, including the role of this type of university in producing agricultural engineers who can innovate with GM to meet regionally-dependent environmental and cultural needs that could increase their sustainability.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2023-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10349173/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10202801","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

Assessment of anti-HIV-1 guide RNA efficacy in cells containing the viral target sequence, corresponding gRNA, and CRISPR/Cas9. 在含有病毒靶序列、相应gRNA和CRISPR/Cas9的细胞中评估抗HIV-1引导RNA的效力。

IF 4.9

Frontiers in genome editing Pub Date : 2023-04-13 eCollection Date: 2023-01-01 DOI: 10.3389/fgeed.2023.1101483

Alexander G Allen, Cheng-Han Chung, Stephen D Worrell, Glad Nwaozo, Rebekah Madrid, Anthony R Mele, Will Dampier, Michael R Nonnemacher, Brian Wigdahl

{"title":"Assessment of anti-HIV-1 guide RNA efficacy in cells containing the viral target sequence, corresponding gRNA, and CRISPR/Cas9.","authors":"Alexander G Allen, Cheng-Han Chung, Stephen D Worrell, Glad Nwaozo, Rebekah Madrid, Anthony R Mele, Will Dampier, Michael R Nonnemacher, Brian Wigdahl","doi":"10.3389/fgeed.2023.1101483","DOIUrl":"10.3389/fgeed.2023.1101483","url":null,"abstract":"The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 gene editing system has been shown to be effective at inhibiting human immunodeficiency virus type 1 (HIV-1). Studies have not consistently used a trackable dual reporter system to determine what cells received the Cas9/gRNA to determine the overall knockdown of HIV. Some studies have used stably transduced cells under drug selection to accomplish this goal. Here a two-color system was used that allows tracking of viral protein expression and which cells received the CRISPR/Cas9 system. These experiments ensured that each gRNA used was a perfect match to the intended target to remove this variable. The data showed that gRNAs targeting the transactivation response element (TAR) region or other highly conserved regions of the HIV-1 genome were effective at stopping viral gene expression, with multiple assays demonstrating greater than 95 percent reduction. Conversely, gRNAs targeting conserved sites of the 5' portion of the U3 region were largely ineffective, demonstrating that the location of edits in the long terminal repeat (LTR) matter with respect to function. In addition, it was observed that a gRNA targeting Tat was effective in a T-cell model of HIV-1 latency. Taken together, these studies demonstrated gRNAs designed to highly conserved functional regions have near 100% efficacy in vitro in cells known to have received the Cas9/gRNA pair.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2023-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10134072/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9393688","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