Frontiers in genome editing最新文献

{"title":"Are null segregants new combinations of heritable material and should they be regulated?","authors":"Jack A Heinemann,&nbsp;Katrin Clark,&nbsp;Tessa C Hiscox,&nbsp;Andrew W McCabe,&nbsp;Sarah Z Agapito-Tenfen","doi":"10.3389/fgeed.2022.1064103","DOIUrl":"https://doi.org/10.3389/fgeed.2022.1064103","url":null,"abstract":"<p><p>Through genome editing and other techniques of gene technology, it is possible to create a class of organism called null segregants. These genetically modified organisms (GMOs) are products of gene technology but are argued to have no lingering vestige of the technology after the segregation of chromosomes or deletion of insertions. From that viewpoint regulations are redundant because any unique potential for the use of gene technology to cause harm has also been removed. We tackle this question of international interest by reviewing the early history of the purpose of gene technology regulation. The active ingredients of techniques used for guided mutagenesis, e.g., site-directed nucleases, such as CRISPR/Cas, are promoted for having a lower potential per reaction to create a hazard. However, others see this as a desirable industrial property of the reagents that will lead to genome editing being used more and nullifying the promised hazard mitigation. The contest between views revolves around whether regulations could alter the risks in the responsible use of gene technology. We conclude that gene technology, even when used to make null segregants, has characteristics that make regulation a reasonable option for mitigating potential harm. Those characteristics are that it allows people to create more harm faster, even if it creates benefits as well; the potential for harm increases with increased use of the technique, but safety does not; and regulations can control harm scaling.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"1064103"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9871356/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10628585","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}

{"title":"Development of Therapeutic RNA Manipulation for Muscular Dystrophy.","authors":"Saifullah,&nbsp;Norio Motohashi,&nbsp;Toshifumi Tsukahara,&nbsp;Yoshitsugu Aoki","doi":"10.3389/fgeed.2022.863651","DOIUrl":"https://doi.org/10.3389/fgeed.2022.863651","url":null,"abstract":"<p><p>Approval of therapeutic RNA molecules, including RNA vaccines, has paved the way for next-generation treatment strategies for various diseases. Oligonucleotide-based therapeutics hold particular promise for treating incurable muscular dystrophies, including Duchenne muscular dystrophy (DMD). DMD is a severe monogenic disease triggered by deletions, duplications, or point mutations in the <i>DMD</i> gene, which encodes a membrane-linked cytoskeletal protein to protect muscle fibers from contraction-induced injury. Patients with DMD inevitably succumb to muscle degeneration and atrophy early in life, leading to premature death from cardiac and respiratory failure. Thus far, the disease has thwarted all curative strategies. Transcriptomic manipulation, employing exon skipping using antisense oligonucleotides (ASO), has made significant progress in the search for DMD therapeutics. Several exon-skipping drugs employing RNA manipulation technology have been approved by regulatory agencies and have shown promise in clinical trials. This review summarizes recent scientific and clinical progress of ASO and other novel RNA manipulations, including RNA-based editing using MS2 coat protein-conjugated adenosine deaminase acting on the RNA (MCP-ADAR) system illustrating the efficacy and limitations of therapies to restore dystrophin. Perhaps lessons from this review will encourage the application of RNA-editing therapy to other neuromuscular disorders.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"863651"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9127466/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10618432","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}

{"title":"Towards social acceptability of genome-edited plants in industrialised countries? Emerging evidence from Europe, United States, Canada, Australia, New Zealand, and Japan.","authors":"Armin Spök,&nbsp;Thorben Sprink,&nbsp;Andrew C Allan,&nbsp;Tomiko Yamaguchi,&nbsp;Christian Dayé","doi":"10.3389/fgeed.2022.899331","DOIUrl":"https://doi.org/10.3389/fgeed.2022.899331","url":null,"abstract":"<p><p>The agricultural biotechnology world has been divided into two blocks; countries adopting GM crops for commercial cultivation (adopters) and others without any or without relevant cultivation of such crops (non-adopters). Meanwhile, an increasing number of adopter countries have exempted certain genome-edited (GE) crops from legal GMO pre-market approval and labelling requirements. Among them are major exporters of agricultural commodities such as United States, Canada, and Australia. Due to the relaxed legislation more GE plants are expected to enter the market soon. Many countries in the non-adopter group, however, depend on import of large volumes of agricultural commodities from adopter countries. Unlike first generation GM, certain GE crops cannot be identified as unambiguously originating from genome editing using available techniques. Consequently, pressure is mounting on non-adopter jurisdictions to reconsider their policies and legislations. Against this backdrop, the paper explores recent developments relevant for social acceptability in selected non-adopters, Japan, New Zealand, the EU, Norway, and Switzerland in contrast to United States, Canada, and Australia. While Japan is already opening-up and Norway and Switzerland are discussing revisions of their policies, the EU and New Zealand are struggling with challenges resulting from high court decisions. In an attempt to take a closer look into the inner dynamics of these developments, the concept of social acceptability proposed by Wüstenhagen et al. (Energy Policy, 2007, 35(5), 2683-2691) is employed. This aids the understanding of developments in the jurisdictions considered and identifies specific or cross-cutting challenges.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"899331"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9473316/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10274274","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}

{"title":"Efficient and error-free correction of sickle mutation in human erythroid cells using prime editor-2.","authors":"Anila George,&nbsp;Nithin Sam Ravi,&nbsp;Kirti Prasad,&nbsp;Lokesh Panigrahi,&nbsp;Sanya Koikkara,&nbsp;Vignesh Rajendiran,&nbsp;Nivedhitha Devaraju,&nbsp;Joshua Paul,&nbsp;Aswin Anand Pai,&nbsp;Yukio Nakamura,&nbsp;Ryo Kurita,&nbsp;Poonkuzhali Balasubramanian,&nbsp;Saravanabhavan Thangavel,&nbsp;Srujan Marepally,&nbsp;Shaji R Velayudhan,&nbsp;Alok Srivastava,&nbsp;Kumarasamypet M Mohankumar","doi":"10.3389/fgeed.2022.1085111","DOIUrl":"https://doi.org/10.3389/fgeed.2022.1085111","url":null,"abstract":"<p><p>Sickle cell anaemia (SCA) is one of the common autosomal recessive monogenic disorders, caused by a transverse point mutation (GAG > GTG) at the sixth codon of the beta-globin gene, which results in haemolytic anaemia due to the fragile RBCs. Recent progress in genome editing has gained attention for the therapeutic cure for SCA. Direct correction of SCA mutation by homology-directed repair relies on a double-strand break (DSB) at the target site and carries the risk of generating beta-thalassaemic mutations if the editing is not error-free. On the other hand, base editors cannot correct the pathogenic SCA mutation resulting from A > T base transversion. Prime editor (PE), the recently described CRISPR/Cas 9 based gene editing tool that enables precise gene manipulations without DSB and unintended nucleotide changes, is a viable approach for the treatment of SCA. However, the major limitation with the use of prime editing is the lower efficiency especially in human erythroid cell lines and primary cells. To overcome these limitations, we developed a modular lenti-viral based prime editor system and demonstrated its use for the precise modelling of SCA mutation and its subsequent correction in human erythroid cell lines. We achieved highly efficient installation of SCA mutation (up to 72%) and its subsequent correction in human erythroid cells. For the first time, we demonstrated the functional restoration of adult haemoglobin without any unintended nucleotide changes or indel formations using the PE2 system. We also validated that the off-target effects mediated by the PE2 system is very minimal even with very efficient on-target conversion, making it a safe therapeutic option. Taken together, the modular lenti-viral prime editor system developed in this study not only expands the range of cell lines targetable by prime editor but also improves the efficiency considerably, enabling the use of prime editor for myriad molecular, genetic, and translational studies.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"1085111"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9808041/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10488550","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}

{"title":"CRISPR medicine for blood disorders: Progress and challenges in delivery.","authors":"Tahereh Mohammadian Gol,&nbsp;Guillermo Ureña-Bailén,&nbsp;Yujuan Hou,&nbsp;Ralph Sinn,&nbsp;Justin S Antony,&nbsp;Rupert Handgretinger,&nbsp;Markus Mezger","doi":"10.3389/fgeed.2022.1037290","DOIUrl":"https://doi.org/10.3389/fgeed.2022.1037290","url":null,"abstract":"<p><p>Blood disorders are a group of diseases including hematological neoplasms, clotting disorders and orphan immune deficiency diseases that affects human health. Current improvements in genome editing based therapeutics demonstrated preclinical and clinical proof to treat different blood disorders. Genome editing components such as Cas nucleases, guide RNAs and base editors are supplied in the form of either a plasmid, an mRNA, or a ribonucleoprotein complex. The most common delivery vehicles for such components include viral vectors (e.g., AAVs and RV), non-viral vectors (e.g., LNPs and polymers) and physical delivery methods (e.g., electroporation and microinjection). Each of the delivery vehicles specified above has its own advantages and disadvantages and the development of a safe transferring method for <i>ex vivo</i> and <i>in vivo</i> application of genome editing components is still a big challenge. Moreover, the delivery of genome editing payload to the target blood cells possess key challenges to provide a possible cure for patients with inherited monogenic blood diseases and hematological neoplastic tumors. Here, we critically review and summarize the progress and challenges related to the delivery of genome editing elements to relevant blood cells in an <i>ex vivo</i> or <i>in vivo</i> setting. In addition, we have attempted to provide a future clinical perspective of genome editing to treat blood disorders with possible clinical grade improvements in delivery methods.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"1037290"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9853164/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9083720","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}

{"title":"Gene Therapy: The Next-Generation Therapeutics and Their Delivery Approaches for Neurological Disorders.","authors":"Abhik Paul,&nbsp;Michael G Collins,&nbsp;Hye Young Lee","doi":"10.3389/fgeed.2022.899209","DOIUrl":"https://doi.org/10.3389/fgeed.2022.899209","url":null,"abstract":"<p><p>Neurological conditions like neurodevelopmental disorders and neurodegenerative diseases are quite complex and often exceedingly difficult for patients. Most of these conditions are due to a mutation in a critical gene. There is no cure for the majority of these neurological conditions and the availability of disease-modifying therapeutics is quite rare. The lion's share of the treatments that are available only provide symptomatic relief, as such, we are in desperate need of an effective therapeutic strategy for these conditions. Considering the current drug development landscape, gene therapy is giving us hope as one such effective therapeutic strategy. Consistent efforts have been made to develop gene therapy strategies using viral and non-viral vectors of gene delivery. Here, we have discussed both of these delivery methods and their properties. We have summarized the relative advantages and drawbacks of viral and non-viral vectors from the perspectives of safety, efficiency, and productivity. Recent developments such as clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated gene editing and its use <i>in vivo</i> have been described here as well. Given recent advancements, gene therapy shows great promise to emerge as a next-generation therapeutic for many of the neurodevelopmental and neurodegenerative conditions.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"4 ","pages":"899209"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9272754/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10352410","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}

{"title":"Genome Editing: Current Technology Advances and Applications for Crop Improvement","authors":"","doi":"10.1007/978-3-031-08072-2","DOIUrl":"https://doi.org/10.1007/978-3-031-08072-2","url":null,"abstract":"","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"282 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72714452","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}

{"title":"Establishment of an Efficient Protoplast Regeneration and Transfection Protocol for Field Cress (<i>Lepidium campestre</i>).","authors":"Sjur Sandgrind,&nbsp;Xueyuan Li,&nbsp;Emelie Ivarson,&nbsp;Annelie Ahlman,&nbsp;Li-Hua Zhu","doi":"10.3389/fgeed.2021.757540","DOIUrl":"https://doi.org/10.3389/fgeed.2021.757540","url":null,"abstract":"<p><p>Field cress (<i>Lepidium campestre</i>) is a potential oilseed crop that has been under domestication in recent decades. CRISPR/Cas9 is a powerful tool for rapid trait improvement and gene characterization and for generating transgene-free mutants using protoplast transfection system. However, protoplast regeneration remains challenging for many plant species. Here we report an efficient protoplast regeneration and transfection protocol for field cress. Important factors such as type of basal media, type/combination of plant growth regulators, and culture duration on different media were optimized. Among the basal media tested, Nitsch was the best for protoplast growth in MI and MII media. For cell wall formation during the early stage of protoplast growth, relatively high auxin concentrations (0.5 mg L^-1 NAA and 2,4-D), without addition of cytokinin was preferred for maintaining protoplast viability. After cell wall formation, 1.1 mg L^-1 TDZ combined with either 0.05 mg L^-1 NAA or 2,4-D was found to efficiently promote protoplast growth. On solid shoot induction medium, 1.1 mg L^-1 TDZ without any auxin resulted in over 80% shoot generation frequency. A longer culture duration in MI medium would inhibit protoplast growth, while a longer culture duration in MII medium significantly delayed shoot formation. Using this optimized protoplast regeneration protocol, we have established an efficient PEG-mediated transfection protocol using a vector harboring the <i>GFP</i> gene, with transfection efficiencies of 50-80%. This efficient protoplast protocol would facilitate further genetic improvement of field cress via genome editing, and be beneficial to development of protoplast regeneration protocols for related plant species.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"3 ","pages":"757540"},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8635052/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9639522","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}

{"title":"Off-Target Analysis in Gene Editing and Applications for Clinical Translation of CRISPR/Cas9 in HIV-1 Therapy.","authors":"Andrew Atkins,&nbsp;Cheng-Han Chung,&nbsp;Alexander G Allen,&nbsp;Will Dampier,&nbsp;Theodore E Gurrola,&nbsp;Ilker K Sariyer,&nbsp;Michael R Nonnemacher,&nbsp;Brian Wigdahl","doi":"10.3389/fgeed.2021.673022","DOIUrl":"https://doi.org/10.3389/fgeed.2021.673022","url":null,"abstract":"<p><p>As genome-editing nucleases move toward broader clinical applications, the need to define the limits of their specificity and efficiency increases. A variety of approaches for nuclease cleavage detection have been developed, allowing a full-genome survey of the targeting landscape and the detection of a variety of repair outcomes for nuclease-induced double-strand breaks. Each approach has advantages and disadvantages relating to the means of target-site capture, target enrichment mechanism, cellular environment, false discovery, and validation of bona fide off-target cleavage sites in cells. This review examines the strengths, limitations, and origins of the different classes of off-target cleavage detection systems including anchored primer enrichment (GUIDE-seq), <i>in situ</i> detection (BLISS), <i>in vitro</i> selection libraries (CIRCLE-seq), chromatin immunoprecipitation (ChIP) (DISCOVER-Seq), translocation sequencing (LAM PCR HTGTS), and <i>in vitro</i> genomic DNA digestion (Digenome-seq and SITE-Seq). Emphasis is placed on the specific modifications that give rise to the enhanced performance of contemporary techniques over their predecessors and the comparative performance of techniques for different applications. The clinical relevance of these techniques is discussed in the context of assessing the safety of novel CRISPR/Cas9 HIV-1 curative strategies. With the recent success of HIV-1 and SIV-1 viral suppression in humanized mice and non-human primates, respectively, using CRISPR/Cas9, rigorous exploration of potential off-target effects is of critical importance. Such analyses would benefit from the application of the techniques discussed in this review.</p>","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"3 ","pages":"673022"},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8525399/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10720299","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}

{"title":"Base editing","authors":"K. Musunuru","doi":"10.1016/b978-0-12-823484-6.00009-8","DOIUrl":"https://doi.org/10.1016/b978-0-12-823484-6.00009-8","url":null,"abstract":"","PeriodicalId":73086,"journal":{"name":"Frontiers in genome editing","volume":"19 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88686880","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}