Frontiers in genome editing最新文献_第8页

Efficient hyperactive piggyBac transgenesis in Plodia pantry moths. Plodia 盘蛾的高效超活性 piggyBac 转基因。

IF 4.9

Frontiers in genome editing Pub Date : 2022-12-23 eCollection Date: 2022-01-01 DOI: 10.3389/fgeed.2022.1074888

Christa Heryanto, Anyi Mazo-Vargas, Arnaud Martin

{"title":"Efficient hyperactive piggyBac transgenesis in Plodia pantry moths.","authors":"Christa Heryanto, Anyi Mazo-Vargas, Arnaud Martin","doi":"10.3389/fgeed.2022.1074888","DOIUrl":"10.3389/fgeed.2022.1074888","url":null,"abstract":"While piggyBac transposon-based transgenesis is widely used in various emerging model organisms, its relatively low transposition rate in butterflies and moths has hindered its use for routine genetic transformation in Lepidoptera. Here, we tested the suitability of a codon-optimized hyperactive piggyBac transposase (hyPBase) in mRNA form to deliver and integrate transgenic cassettes into the genome of the pantry moth Plodia interpunctella. Co-injection of hyPBase mRNA with donor plasmids successfully integrated 1.5-4.4 kb expression cassettes driving the fluorescent markers EGFP, DsRed, or EYFP in eyes and glia with the 3xP3 promoter. Somatic integration and expression of the transgene in the G0 injected generation was detectable from 72-h embryos and onward in larvae, pupae and adults carrying a recessive white-eyed mutation. Overall, 2.5% of injected eggs survived into transgene-bearing adults with mosaic fluorescence. Subsequent outcrossing of fluorescent G0 founders transmitted single-insertion copies of 3xP3::EGFP and 3xP3::EYFP and generated stable isogenic lines. Random in-crossing of a small cohort of G0 founders expressing 3xP3::DsRed yielded a stable transgenic line segregating for more than one transgene insertion site. We discuss how hyPBase can be used to generate stable transgenic resources in Plodia and other moths.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"1074888"},"PeriodicalIF":4.9,"publicationDate":"2022-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9816379/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10874972","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

Imaging tools for plant nanobiotechnology. 植物纳米生物技术的成像工具。

IF 4.9

Frontiers in genome editing Pub Date : 2022-12-08 eCollection Date: 2022-01-01 DOI: 10.3389/fgeed.2022.1029944

Bin Zhao, Zhongxu Luo, Honglu Zhang, Huan Zhang

{"title":"Imaging tools for plant nanobiotechnology.","authors":"Bin Zhao, Zhongxu Luo, Honglu Zhang, Huan Zhang","doi":"10.3389/fgeed.2022.1029944","DOIUrl":"10.3389/fgeed.2022.1029944","url":null,"abstract":"The successful application of nanobiotechnology in biomedicine has greatly changed the traditional way of diagnosis and treating of disease, and is promising for revolutionizing the traditional plant nanobiotechnology. Over the past few years, nanobiotechnology has increasingly expanded into plant research area. Nanomaterials can be designed as vectors for targeted delivery and controlled release of fertilizers, pesticides, herbicides, nucleotides, proteins, etc. Interestingly, nanomaterials with unique physical and chemical properties can directly affect plant growth and development; improve plant resistance to disease and stress; design as sensors in plant biology; and even be used for plant genetic engineering. Similarly, there have been concerns about the potential biological toxicity of nanomaterials. Selecting appropriate characterization methods will help understand how nanomaterials interact with plants and promote advances in plant nanobiotechnology. However, there are relatively few reviews of tools for characterizing nanomaterials in plant nanobiotechnology. In this review, we present relevant imaging tools that have been used in plant nanobiotechnology to monitor nanomaterial migration, interaction with and internalization into plants at three-dimensional lengths. Including: 1) Migration of nanomaterial into plant organs 2) Penetration of nanomaterial into plant tissues (iii)Internalization of nanomaterials by plant cells and interactions with plant subcellular structures. We compare the advantages and disadvantages of current characterization tools and propose future optimal characterization methods for plant nanobiotechnology.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"1029944"},"PeriodicalIF":4.9,"publicationDate":"2022-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9772283/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10436908","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 nuclease off-target activity and mitigation strategies. CRISPR 核酸酶脱靶活性及缓解策略。

IF 4.9

Frontiers in genome editing Pub Date : 2022-11-10 eCollection Date: 2022-01-01 DOI: 10.3389/fgeed.2022.1050507

Beeke Wienert, M Kyle Cromer

{"title":"CRISPR nuclease off-target activity and mitigation strategies.","authors":"Beeke Wienert, M Kyle Cromer","doi":"10.3389/fgeed.2022.1050507","DOIUrl":"10.3389/fgeed.2022.1050507","url":null,"abstract":"The discovery of CRISPR has allowed site-specific genomic modification to become a reality and this technology is now being applied in a number of human clinical trials. While this technology has demonstrated impressive efficacy in the clinic to date, there remains the potential for unintended on- and off-target effects of CRISPR nuclease activity. A variety of in silico-based prediction tools and empirically derived experimental methods have been developed to identify the most common unintended effect-small insertions and deletions at genomic sites with homology to the guide RNA. However, large-scale aberrations have recently been reported such as translocations, inversions, deletions, and even chromothripsis. These are more difficult to detect using current workflows indicating a major unmet need in the field. In this review we summarize potential sequencing-based solutions that may be able to detect these large-scale effects even at low frequencies of occurrence. In addition, many of the current clinical trials using CRISPR involve ex vivo isolation of a patient's own stem cells, modification, and re-transplantation. However, there is growing interest in direct, in vivo delivery of genome editing tools. While this strategy has the potential to address disease in cell types that are not amenable to ex vivo manipulation, in vivo editing has only one desired outcome-on-target editing in the cell type of interest. CRISPR activity in unintended cell types (both on- and off-target) is therefore a major safety as well as ethical concern in tissues that could enable germline transmission. In this review, we have summarized the strengths and weaknesses of current editing and delivery tools and potential improvements to off-target and off-tissue CRISPR activity detection. We have also outlined potential mitigation strategies that will ensure that the safety of CRISPR keeps pace with efficacy, a necessary requirement if this technology is to realize its full translational potential.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"1050507"},"PeriodicalIF":4.9,"publicationDate":"2022-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9685173/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10658011","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

Plant biomacromolecule delivery methods in the 21st century. 21 世纪的植物生物大分子输送方法。

IF 4.9

Frontiers in genome editing Pub Date : 2022-10-14 eCollection Date: 2022-01-01 DOI: 10.3389/fgeed.2022.1011934

Sachin Rustgi, Salman Naveed, Jonathan Windham, Huan Zhang, Gözde S Demirer

引用次数: 0

Enabling Genome Editing for Enhanced Agricultural Sustainability 实现基因组编辑以增强农业可持续性

Frontiers in genome editing Pub Date : 2022-05-18 DOI: 10.3389/fgeed.2022.898950

F. Keiper, A. Atanassova

引用次数: 2

Advantages and Limitations of Gene Therapy and Gene Editing for Friedreich's Ataxia. 弗里德里希共济失调的基因治疗和基因编辑的优势和局限性。

IF 5.4

Frontiers in genome editing Pub Date : 2022-05-17 eCollection Date: 2022-01-01 DOI: 10.3389/fgeed.2022.903139

Anusha Sivakumar, Stephanie Cherqui

{"title":"Advantages and Limitations of Gene Therapy and Gene Editing for Friedreich's Ataxia.","authors":"Anusha Sivakumar, Stephanie Cherqui","doi":"10.3389/fgeed.2022.903139","DOIUrl":"10.3389/fgeed.2022.903139","url":null,"abstract":"Friedreich's ataxia (FRDA) is an inherited, multisystemic disorder predominantly caused by GAA hyper expansion in intron 1 of frataxin (FXN) gene. This expansion mutation transcriptionally represses FXN, a mitochondrial protein that is required for iron metabolism and mitochondrial homeostasis, leading to neurodegerative and cardiac dysfunction. Current therapeutic options for FRDA are focused on improving mitochondrial function and increasing frataxin expression through pharmacological interventions but are not effective in delaying or preventing the neurodegeneration in clinical trials. Recent research on in vivo and ex vivo gene therapy methods in FRDA animal and cell models showcase its promise as a one-time therapy for FRDA. In this review, we provide an overview on the current and emerging prospects of gene therapy for FRDA, with specific focus on advantages of CRISPR/Cas9-mediated gene editing of FXN as a viable option to restore endogenous frataxin expression. We also assess the potential of ex vivo gene editing in hematopoietic stem and progenitor cells as a potential autologous transplantation therapeutic option and discuss its advantages in tackling FRDA-specific safety aspects for clinical translation.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"903139"},"PeriodicalIF":5.4,"publicationDate":"2022-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9157421/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41241787","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 Editing in Peanut Using CRISPR/nCas9 利用CRISPR/nCas9对花生进行碱基编辑

Frontiers in genome editing Pub Date : 2022-05-12 DOI: 10.3389/fgeed.2022.901444

A. Neelakandan, B. Subedi, S. Traore, P. Binagwa, D. Wright, G. He

{"title":"Base Editing in Peanut Using CRISPR/nCas9","authors":"A. Neelakandan, B. Subedi, S. Traore, P. Binagwa, D. Wright, G. He","doi":"10.3389/fgeed.2022.901444","DOIUrl":"https://doi.org/10.3389/fgeed.2022.901444","url":null,"abstract":"Peanut (Arachis hypogaea L.), an allotetraploid legume of the Fabaceae family, is able to thrive in tropical and subtropical regions and is considered as a promising oil seed crop worldwide. Increasing the content of oleic acid has become one of the major goals in peanut breeding because of health benefits such as reduced blood cholesterol level, antioxidant properties and industrial benefits such as longer shelf life. Genomic sequencing of peanut has provided evidence of homeologous AhFAD2A and AhFAD2B genes encoding Fatty Acid Desaturase2 (FAD2), which are responsible for catalyzing the conversion of monounsaturated oleic acid into polyunsaturated linoleic acid. Research studies demonstrate that mutations resulting in a frameshift or stop codon in an FAD2 gene leads to higher oleic acid content in oil. In this study, two expression vectors, pDW3873 and pDW3876, were constructed using Cas9 fused to different deaminases, which were tested as tools to induce point mutations in the promoter and the coding sequences of peanut AhFAD2 genes. Both constructs harbor the single nuclease null variant, nCas9 D10A, to which the PmCDA1 cytosine deaminase was fused to the C-terminal (pDW3873) while rAPOBEC1 deaminase and an uracil glycosylase inhibitor (UGI) were fused to the N-terminal and the C-terminal respectively (pDW3876). Three gRNAs were cloned independently into both constructs and the functionality and efficiency were tested at three target sites in the AhFAD2 genes. Both constructs displayed base editing activity in which cytosine was replaced by thymine or other bases in the targeted editing window. pDW3873 showed higher efficiency compared to pDW3876 suggesting that the former is a better base editor in peanut. This is an important step forward considering introgression of existing mutations into elite varieties can take up to 15 years making this tool a benefit for peanut breeders, farmers, industry and ultimately for consumers.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43872135","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

Potential of Genome Editing to Capture Diversity From Australian Wild Rice Relatives 基因组编辑获取澳大利亚野生稻近缘品种多样性的潜力

Frontiers in genome editing Pub Date : 2022-04-27 DOI: 10.3389/fgeed.2022.875243

Muhammad Abdullah, P. Okemo, A. Furtado, R. Henry

{"title":"Potential of Genome Editing to Capture Diversity From Australian Wild Rice Relatives","authors":"Muhammad Abdullah, P. Okemo, A. Furtado, R. Henry","doi":"10.3389/fgeed.2022.875243","DOIUrl":"https://doi.org/10.3389/fgeed.2022.875243","url":null,"abstract":"Rice, a staple food worldwide and a model crop, could benefit from the introduction of novel genetics from wild relatives. Wild rice in the AA genome group closely related to domesticated rice is found across the tropical world. Due to their locality outside the range of domesticated rice, Australian wild rice populations are a potential source of unique traits for rice breeding. These rice species provide a diverse gene pool for improvement that could be utilized for desirable traits such as stress resistance, disease tolerance, and nutritional qualities. However, they remain poorly characterized. The CRISPR/Cas system has revolutionized gene editing and has improved our understanding of gene functions. Coupled with the increasing availability of genomic information on the species, genes in Australian wild rice could be modified through genome editing technologies to produce new domesticates. Alternatively, beneficial alleles from these rice species could be incorporated into cultivated rice to improve critical traits. Here, we summarize the beneficial traits in Australian wild rice, the available genomic information and the potential of gene editing to discover and understand the functions of novel alleles. Moreover, we discuss the potential domestication of these wild rice species for health and economic benefits to rice production globally.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48479248","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

Spatiotemporal Regulation of CRISPR/Cas9 Enables Efficient, Precise, and Heritable Edits in Plant Genomes CRISPR/Cas9的时空调控使植物基因组高效、精确和可遗传编辑成为可能

Frontiers in genome editing Pub Date : 2022-04-26 DOI: 10.3389/fgeed.2022.870108

Farhanur Rahman, Apurva Mishra, Archit Gupta, Rita Sharma

引用次数: 4

CRISPR-Mediated Activation of αV Integrin Subtypes Promotes Neuronal Differentiation of Neuroblastoma Neuro2a Cells crispr介导的αV整合素亚型激活促进神经母细胞瘤Neuro2a细胞的神经元分化

Frontiers in genome editing Pub Date : 2022-04-12 DOI: 10.3389/fgeed.2022.846669

Sara Riccardi, L. Cingolani, F. Jaudon

{"title":"CRISPR-Mediated Activation of αV Integrin Subtypes Promotes Neuronal Differentiation of Neuroblastoma Neuro2a Cells","authors":"Sara Riccardi, L. Cingolani, F. Jaudon","doi":"10.3389/fgeed.2022.846669","DOIUrl":"https://doi.org/10.3389/fgeed.2022.846669","url":null,"abstract":"Neuronal differentiation is a complex process whose dysfunction can lead to brain disorders. The development of new tools to target specific steps in the neuronal differentiation process is of paramount importance for a better understanding of the molecular mechanisms involved, and ultimately for developing effective therapeutic strategies for neurodevelopmental disorders. Through their interactions with extracellular matrix proteins, the cell adhesion molecules of the integrin family play essential roles in the formation of functional neuronal circuits by regulating cell migration, neurite outgrowth, dendritic spine formation and synaptic plasticity. However, how different integrin receptors contribute to the successive phases of neuronal differentiation remains to be elucidated. Here, we implemented a CRISPR activation system to enhance the endogenous expression of specific integrin subunits in an in vitro model of neuronal differentiation, the murine neuroblastoma Neuro2a cell line. By combining CRISPR activation with morphological and RT-qPCR analyses, we show that integrins of the αV family are powerful inducers of neuronal differentiation. Further, we identify a subtype-specific role for αV integrins in controlling neurite outgrowth. While αVβ3 integrin initiates neuronal differentiation of Neuro2a cells under proliferative conditions, αVβ5 integrin appears responsible for promoting a complex arborization in cells already committed to differentiation. Interestingly, primary neurons exhibit a complementary expression pattern for β3 and β5 integrin subunits during development. Our findings reveal the existence of a developmental switch between αV integrin subtypes during differentiation and suggest that a timely controlled modulation of the expression of αV integrins by CRISPRa provides a means to promote neuronal differentiation.","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44371044","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: 4