Editing Technology Research Articles

LGG-07. Novel CRISPR/Cas9 induced KIAA1549:BRAF fusion model for preclinical studies of pediatric gliomas

BACKGROUND: Pediatric Low Grade Gliomas (pLGG) are the most common group of central nervous system (CNS) tumors in children and cause significant morbidities. Pilocytic astrocytoma (PA) is the most frequent pLGG. KIAA1549:BRAF fusion is a well-established oncogenic driver in PA. Oncogene induced senescence (OIS) has prevented establishing PA cultures for in vitro and in vivo studies. Here we look at a novel NIH-3T3 fibroblast model harboring KIAA1549:BRAF fusion gene via CRISPR/Cas9 somatic genome engineering technology. OBJECTIVES: Establishing that the CRISPR/Cas9 edited NIH-3T3 fibroblast model with the KIAA1549:BRAF fusion is valuable for in vitro and in vivo studies without early OIS. DESIGN/METHOD: CRISPR/Cas9 editing technology was used to establish a KIAA1549:BRAF fusion positive cell model. This cell model was studied in vitro with MEK inhibitor cobimetinib (GDC-0973) and using WST-1 viability assay, clonogenic assay, senescence β-galactosidase staining, and western blot. In vivo murine models with subcutaneous fusion positive NIH-3T3 fibroblast tumors were treated with GDC-0973. Survival studies and tissue studies were subsequently done. RESULTS: A fusion positive NIH-3T3 fibroblast model was successfully established. Increased BRAF cDNA expression and higher levels of p-ERK were observed. In vitro studies showed decreased viability with GDC-0973. Clonogenic assay showed qualitative and quantitative decreases in viable cells. P-ERK target inhibition was established without induction of senescence. In vivo studies demonstrated successful subcutaneous tumor implantation, therapy efficacy, and target inhibition. CONCLUSION: CNS tumors, most commonly pLGG, in children cause significant morbidities. KIAA1549:BRAF fusion is an oncogenic driver in PA. In vitro and in vivo studies are important for pre-clinical models. OIS has prevented establishing adequate fusion positive animal cell models. Here we have demonstrated a successful CRISPR/Cas9 edited fusion positive NIH-3T3 fibroblast model.

Analysis and Evaluation on the Harmfulness of Sports Dance Based on Intelligent Computing

With the rapid development of motion analysis and editing technology, more and more animation technologies based on real human motion analysis are widely used in games, medicine, film and television, sports, and other fields. The role of this technology in the teaching system of sports dance arrangement cannot be ignored. This paper studies the application of intelligent algorithm in injury analysis of sports dance choreography and puts forward a smooth and natural motion editing method which is more in line with the characteristics of human motion. The square spherical surface intelligent algorithm is used to naturally transition the rotation posture of each frame of human joint, and the action modification of computer-aided dance choreography system is more in line with the real action of human body. Simulation results show that interframe data can be used for the same type of dance. Different types of dance arrangement can adopt the method of inserting a new framework between the two dance movements, so as to finally meet the needs of sports dance coaches and students to observe their own dance movements in an all-round way, so as to facilitate the arrangement and modification of sports dance movement design. It provides a certain reference for the application and development of virtual technology in the field of sports and greatly improves the current situation of sports dance teaching equipment. Through animation simulation technology, we can effectively avoid the injury caused by dangerous movements, provide a way of observation and improvement for coaches’ dance teaching, and effectively improve the training effect. Compared with traditional dance choreography methods, the intelligent algorithm in this study can make the animation display of sports dance more coherent and realistic and effectively solve the needs of current events observation and modification in sports dance choreography. It can produce complex movements with simple movements and help teachers arrange and teach sports dance. Avoid the accidental injury caused by the limitation of time and space in traditional teaching and realize the modernization and intelligence of physical education teaching.

Seamless Gene Correction in the Human Cystic Fibrosis Transmembrane Conductance Regulator Locus by Vector Replacement and Vector Insertion Events.

Background: Gene correction via homology directed repair (HDR) in patient-derived induced pluripotent stem (iPS) cells for regenerative medicine are becoming a more realistic approach to develop personalized and mutation-specific therapeutic strategies due to current developments in gene editing and iPSC technology. Cystic fibrosis (CF) is the most common inherited disease in the Caucasian population, caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. Since CF causes significant multi-organ damage and with over 2,000 reported CFTR mutations, CF patients could be one prominent population benefiting from gene and cell therapies. When considering gene-editing techniques for clinical applications, seamless gene corrections of the responsible mutations, restoring native “wildtype” DNA sequence without remnants of drug selectable markers or unwanted DNA sequence changes, would be the most desirable approach. Result: The studies reported here describe the seamless correction of the W1282X CFTR mutation using CRISPR/Cas9 nickases (Cas9n) in iPS cells derived from a CF patient homozygous for the W1282X Class I CFTR mutation. In addition to the expected HDR vector replacement product, we discovered another class of HDR products resulting from vector insertion events that created partial duplications of the CFTR exon 23 region. These vector insertion events were removed via intrachromosomal homologous recombination (IHR) enhanced by double nicking with CRISPR/Cas9n which resulted in the seamless correction of CFTR exon 23 in CF-iPS cells. Conclusion: We show here the removal of the drug resistance cassette and generation of seamless gene corrected cell lines by two independent processes: by treatment with the PiggyBac (PB) transposase in vector replacements or by IHR between the tandemly duplicated CFTR gene sequences.

Plant Bioactive Compounds as an Intrinsic and Sustainable Tool to Enhance the Microbial Safety of Crops.

Outbreaks of produce-associated foodborne illness continue to pose a threat to human health worldwide. New approaches are necessary to improve produce safety. Plant innate immunity has potential as a host-based strategy for the deactivation of enteric pathogens. In response to various biotic and abiotic threats, plants mount defense responses that are governed by signaling pathways. Once activated, these result in the release of reactive oxygen and nitrogen species in addition to secondary metabolites that aim at tempering microbial infection and pest attack. These phytochemicals have been investigated as alternatives to chemical sanitization, as many are effective antimicrobial compounds in vitro. Their antagonistic activity toward enteric pathogens may also provide an intrinsic hurdle to their viability and multiplication in planta. Plants can detect and mount basal defenses against enteric pathogens. Evidence supports the role of plant bioactive compounds in the physiology of Salmonella enterica, Escherichia coli, and Listeria monocytogenes as well as their fitness on plants. Here, we review the current state of knowledge of the effect of phytochemicals on enteric pathogens and their colonization of plants. Further understanding of the interplay between foodborne pathogens and the chemical environment on/in host plants may have lasting impacts on crop management for enhanced microbial safety through translational applications in plant breeding, editing technologies, and defense priming.

A Portrait of Intratumoral Genomic and Transcriptomic Heterogeneity at Single-Cell Level in Colorectal Cancer.

In the study of cancer, omics technologies are supporting the transition from traditional clinical approaches to precision medicine. Intra-tumoral heterogeneity (ITH) is detectable within a single tumor in which cancer cell subpopulations with different genome features coexist in a patient in different tumor areas or may evolve/differ over time. Colorectal carcinoma (CRC) is characterized by heterogeneous features involving genomic, epigenomic, and transcriptomic alterations. The study of ITH is a promising new frontier to lay the foundation towards successful CRC diagnosis and treatment. Genome and transcriptome sequencing together with editing technologies are revolutionizing biomedical research, representing the most promising tools for overcoming unmet clinical and research challenges. Rapid advances in both bulk and single-cell next-generation sequencing (NGS) are identifying primary and metastatic intratumoral genomic and transcriptional heterogeneity. They provide critical insight in the origin and spatiotemporal evolution of genomic clones responsible for early and late therapeutic resistance and relapse. Single-cell technologies can be used to define subpopulations within a known cell type by searching for differential gene expression within the cell population of interest and/or effectively isolating signal from rare cell populations that would not be detectable by other methods. Each single-cell sequencing analysis is driven by clustering of cells based on their differentially expressed genes. Genes that drive clustering can be used as unique markers for a specific cell population. In this review we analyzed, starting from published data, the possible achievement of a transition from clinical CRC research to precision medicine with an emphasis on new single-cell based techniques; at the same time, we focused on all approaches and issues related to this promising technology. This transition might enable noninvasive screening for early diagnosis, individualized prediction of therapeutic response, and discovery of additional novel drug targets.

Ethics in Gene Editing

For thousands of years, humans have felt the need to understand the world around them—and ultimately manipulate it to best serve their needs. There are always ethical questions to address, especially when the manipulation involves the human genome. There is currently an urgent need to actively pursue those conversations as com- mercial gene sequencing and editing technologies have become more accessible and affordable. Gene Editing has enormous potential both as a research tool and a therapeutic intervention. While other types of gene editing are relatively uncontroversial, gene editing has been strongly resisted.

Ethics in Gene Editing

For thousands of years, humans have felt the need to understand the world around them—and ultimately manipulate it to best serve their needs. There are always ethical questions to address, especially when the manipulation involves the human genome. There is currently an urgent need to actively pursue those conversations as com- mercial gene sequencing and editing technologies have become more accessible and affordable. Gene Editing has enormous potential both as a research tool and a therapeutic intervention. While other types of gene editing are relatively uncontroversial, gene editing has been strongly resisted.

Ethics In Gene Editing

For thousands of years, humans have felt the need to understand the world around them—and ultimately manipulate it to best serve their needs. There are always ethical questions to address, especially when the manipulation involves the human genome. There is currently an urgent need to actively pursue those conversations as com- mercial gene sequencing and editing technologies have become more accessible and affordable. Gene Editing has enormous potential both as a research tool and a therapeutic intervention. While other types of gene editing are relatively uncontroversial, gene editing has been strongly resisted.

Genetic engineering and genome editing techniques in peanut plants

Research has long been associated with human life. In the effort to make a living, many experts who have contributed to the modernization of traditional research methods by conducting various research activities. In this process, professionals, from farmers to senior researchers, have done their part by developing plants that can tolerate or resist to disease. The growing population, climate change and plant disease are having a devastating effect on food security. In particular, it is essential to increase food production by producing high yielding crops of good quality, that may ensure food security. Recently, different gene- editing technologies have been developed. These techniques have been applied in many research fields and their development has provided economic benefits to farmers. Agrobacterium-mediated and biolistic methods are very important techniques for transforming genetic materials in plants. Genome- editing technologies are recent and highly applied in plant research to improve genes associated with yield, disease resistance and drought resistance. For example, Zinc-finger Nucleases (ZFNS), Transcription Activator-like Effector Nucleases (TALEN), and Clustered Regularly Interspaced Short Palindromic Repeats system (CRISPR/ Cas9) methods are now widely applied by researchers and are playing a positive role in increasing production and productivity. Of the gene- editing technology, CRISPR/ Cas9 is widely applied in plant breeding programme as it is easy to use and cost-effective. In this review, we mainly focus on peanut plant, which is an important oil-bearing allotetraploid crop. Therefore, peanut gene editing-technology could increase the oleic acid content in edible peanut oil. Thus, genome editing and gene transformation technologies are extensively explored in this review.

Research on Computer Two-dimensional Animation Creation Tools and Processing and Editing Technology

This article analyzes the creation process of two-dimensional computer animation. The research content of this paper includes key frame generation, production of intermediate images, layered production, coloring and rehearsal processing, reasonable use of library images, combined with specific applications of authoring tools such as Flash software, animo software, TOONZ software, and FlipBook software. The author studies the specific application of computer two-dimensional animation processing and editing technology. The purpose of this article is to optimize the process of computer two-dimensional animation creation and improve the application effect of the two-dimensional animation produced.

112 How Will the Purebred Association Adapt to a Changing Beef Industry?

Abstract The role of the breed association has historically been to keep a registry of a pure breed, aggregate the performance data surrounding that one breed, develop breeding and selection tools, and conduct breed promotion. Larger associations have been able to augment that with operating magazines and other media, running branded beef programs, feeder calf marketing programs, and genetic evaluation for other breeds. The relevance of breed associations is being and will continue to be challenged as genomics and large commercial databases develop and allow for breeding and selection tools to be developed independently by large breeders or private entities. Gene editing and other such technology will also challenge the traditional seedstock models and opens the door for proprietary genetic lines. Breed associations may need to modify their traditional policies to incorporate these innovations. Supply chains will continue to become far more sophisticated and will incorporate more genetic information to guide management decisions and potentially validate brand promises around sustainability. To stay relevant, breed associations of the future will need to do the following: Balance the needs of diverse membership (show, hobby, lifestyle, etc.) with commercial industry value and significance. Have access to large amounts of data and be leaders in adopting the most current technologies. Deliver tools for breeders that enhance the profitability of commercial producers – identify optimum production levels vs maximum outputs. Work collaboratively with multiple supply chains providing the needed genetic information. Be a significant educational resource to breeders and commercial producers. Be a leader in research on breed improvement and genetic advancement. Have value-added programs that create real and sustained pull-through demand for the end product.

Childbearing: Re-production or Creation

The assisted reproduction possibilities, enhanced by the technology of gene editing, are considered in the context of the demographic trend to rationalize childbearing, expressed in the formula “a desired child at a desired time”. Applying of gene editing in human embryos illustrates the transition from quantitative measurements of procreation to the choice of the child "quality". The gene editing in the human embryo is presented as a consistent development of technological interventions into the area of human reproduction, it acts as a link in the expansion of implementation of the preimplantation diagnosis (PGD) of embryos method. When in the case of PGD the main purpose was selection, then nowadays it becomes possible to combine the selection of embryos with the modification of the genetic structure. The participation of geneticists in artificial reproduction increases the efficiency of IVF cycles and at the same time enhances the technological prosthetics of the childbearing process. Following the increase of supply in artificial reproduction, it becomes possible not just to plan a pregnancy, but also to control genetic bases that are still beyond human control, but also to determine the characteristics of an unborn child. A new anthropological situation of displacement or substitution of the value aspects of procreation, understood as procreation, i.e. a human creation, by reproduction - the production, the construction of an ability to give birth to a child first and then the children themselves. As a result, the meanings of childbearing are transformed, the reproductive attitudes and parents' intentions change. Arguments in discussions about the admissibility of intervention with the embryonic genome reflect contradictions in understanding the value of natural foundations in human birth, which are presented in procreation and reproduction opposition. An orientation towards the “quality” of children in social terms has its maintain in a transformation of family ties, in a values shift of kinship and parenthood. Thus, the gene-editing of the embryo enhances two tendencies in demographic behaviour: concern for the child “quality”, which entails eugenic consequences, and a constructive and technological trend in reproduction that threatening the natural foundations of procreation.

ТРАНСФОРМАЦІЯ ХУДОЖНІХ ФОРМ УКРАЇНСЬКОЇ ДОКУДРАМИ В КОНТЕКСТІ ТЕНДЕНЦІЙ ПОСТМОДЕРНУ

The purpose of the article is to identify the transformational form-creating processes of the Ukrainian television art of the 21st century on the example of the genre of documentary drama in the context of postmodern trends. Research Methodology. The following methods have been applied in the article: analytical and historical, cultural methods (to understand process of infotainment formation and development in the context of the Ukrainian television space at the beginning of the 21st century), cultural method (to develop the necessary view of the modern Ukrainian television art, the special feature of the postmodern concept implementation in the media space, the issues of mass culture and mass communications), the method of art history, conceptual and cognitive analyses, which contributed to the understanding of the conceptual representation of information and entertainment presentation of material in the modern Ukrainian television. Scientific Novelty. The article presented an attempt to determine the peculiarity of the aesthetics of the Ukrainian documentary drama in the context of infotainment of domestic television, and also to demonstrate its main artistic, figurative and technological features; to identify challenges and prospects for further development of a new genre of the modern television art. Conclusions. In the development of the domestic docudrama, wide possibilities of video editing and post-production technologies are used, but the creators are particularly demanding in choosing the means that are organic to the artistic language of films. The use of video editing and computer visual effects is an expressive tool for creating a “digital chronotope” that can reveal the philosophy of documentary drama and inject it with virtuality. Evolution tendencies of the Ukrainian documentary drama in the process of infotainment of television art give evidence of the establishment of a democratic screen language aimed at a wide audience, and at the same time of the strengthening of the philosophical sound of works. In our opinion, the process of globalization and convergence of techniques and methods of television documentary and infotainment, interaction and interpenetration of various verbal and non-verbal characteristics in the near future will have transition from quantity to quality.

Prohibited Editing and transition technology in single-shot movies, "1917 Film as a model”

انطلاقا من مصطلح (المونتاج الممنوع) الذي اطلقه الناقد الفرنسي (اندرية بازان) Andre Bazin كاسلوب في معالجة الافلام التي تعتمد على (الميزانسين) المتحقق بفعل الكاميرا وامكانيتها في التصوير وتوظيف عمق الميدان كذلك امكانية الحركة الحرة دون توقف في بيئة التصوير، حتى يتسنى الابتعاد عن المونتاج قدر الامكان (المونتاج الذي يفسد التركيز ويشتت الانتباه ويبتعد عن الواقعية التي هي اهم ركائز بازن التنظيرية في التصوير)، فالسعي كان خلف سينما تصور مواضيعها بلقطة واحدة متكاملة بكل تفاصيلها مقتربة بذلك الى الواقعية من دون اي تدخل للمونتاج، من هذا المفهوم انطلقت دراستنا التي تناولت موضوع معالجة مواضيع الافلام المنجزة من غير المونتاج اوالتقطيع الذي يبضع الفيلم والتي يطلق عليها افلام اللقطة الواحدة المستمرة، ومن تلك الافلام هو الفيلم الحربي (1917) الذي انتج في العام نفسه الذي انطلقت منها شرارة وباء كورونا عام 2019. انطلقت دراستنا هذه الى تشخيص كيفية معالجة قصص الافلام الروائية بلقطة طويلة واحدة مستمرة من غير المونتاج، على الرغم من ان تلك الافلام عمليا تشكلت من مجموعة من اللقطات الا ان تكنيكا معينا قام بتركيب تلك اللقطات الطويلة الى بعضها بعض تركيبا حرفيا وليس تعبيريا ليعطي الاحساس ويثير الانتباه الى ان الفيلم مركب من لقطة واحدة متواصلة التدفق، هذا التكنيك تمحور الى انماط معينة من الاليات والتقنيات تسهل عملية تركيب اللقطات على شكل (القطع غير المرئي) والتي تمنح بناء الفيلم مفهوم (الفيلم اللقطة).خرجت الدراسة ببعض النتائج ومنها تحديد اسلوب التكنيك المعتمد في الانتقال والذي ساعد من الاحساس من ان الفيلم يبدو كلقطة واحدة وبوب اسلوب هذا التكنيك الى (الخداع او المكيدة واخفاء الانتقال تحت القناع وحركة الكاميرا الخاطفة والمؤثرات الصورية بكل تشكيلاتها وتكنيك الاظلام و التشابك)

CRISPR-Cas9 System for Plant Genome Editing: Current Approaches and Emerging Developments

Targeted genome editing using CRISPR-Cas9 has been widely adopted as a genetic engineering tool in various biological systems. This editing technology has been in the limelight due to its simplicity and versatility compared to other previously known genome editing platforms. Several modifications of this editing system have been established for adoption in a variety of plants, as well as for its improved efficiency and portability, bringing new opportunities for the development of transgene-free improved varieties of economically important crops. This review presents an overview of CRISPR-Cas9 and its application in plant genome editing. A catalog of the current and emerging approaches for the implementation of the system in plants is also presented with details on the existing gaps and limitations. Strategies for the establishment of the CRISPR-Cas9 molecular construct such as the selection of sgRNAs, PAM compatibility, choice of promoters, vector architecture, and multiplexing approaches are emphasized. Progress in the delivery and transgene detection methods, together with optimization approaches for improved on-target efficiency are also detailed in this review. The information laid out here will provide options useful for the effective and efficient exploitation of the system for plant genome editing and will serve as a baseline for further developments of the system. Future combinations and fine-tuning of the known parameters or factors that contribute to the editing efficiency, fidelity, and portability of CRISPR-Cas9 will indeed open avenues for new technological advancements of the system for targeted gene editing in plants.

Analysis on the Art Skills of Editing Radio and TV News in the Era of Financial Media

<p>Under the background of the new era, network technology, information technology and new media technology have been applied to various fields, effectively promoting social progress and industry development. In particular, the application of big data, cloud computing and multimedia technology in the radio and television industry not only accelerates the speed of information transmission and increases the number of information dissemination, but also truly realizes information sharing and promotes the sustainable development of the radio and television industry. Under the background of the new era, the information collection and processing technology and editing technology in the radio and television news industry have been improved, which also puts forward new requirements for the editing skills of radio and television news. Based on the author's own news practice, through analyzing the editing skills of radio and television news in the new era, the author puts forward some thoughts and countermeasures to provide useful reference for more radio and television news from the north.</p>

Ethical considerations of gene editing and genetic selection.

For thousands of years, humans have felt the need to understand the world around them—and ultimately manipulate it to best serve their needs. There are always ethical questions to address, especially when the manipulation involves the human genome. There is currently an urgent need to actively pursue those conversations as commercial gene sequencing and editing technologies have become more accessible and affordable. This paper explores the ethical considerations of gene editing (specifically germline) and genetic selection—including the hurdles researchers will face in trying to develop new technologies into viable therapeutic options.

CREATING LOW-COST ANIMATION VIDEO USING ONLINE PLATFORM: A LEARNING MEDIA USER REVIEW

Background: A shift in modern learner learning preference and expansive growth of computer capacities leads to a more widely use of multimedia instructions, including animation videos. Creating animation instructions can be time and resource-consuming. A means of low-cost animation video making platform is favorable. This article aims to review an example of online platform for animation video making.Learning Media Review: Powtoonâ is an online video making and editing platform for producing 2D animation videos. It provides animation attributes, i.e. icons, characters, text, background pictures, etc., and editing technology. The features can be accessed in several bundling and pricing alternatives depends on users need. The article also describes some points in terms of advantages and disadvantages of this online platform. Animation in education can be used for gaining learners’ attention; demonstration of a process or procedures; and supporting explanation for conceptual understanding. Brief discussions about educational theories related to animation video development and use are included.Conclusion: Alternatives of online platforms are available and can be used to develop low-cost educational animation video. Teachers as designer should develop a meaningful concept and storyline guided by the application of sound educational theories during the development and in class implementation.

An Analysis of Significant Role of Technology in Education and Potentiality of Digital Tools in Enhancing Literacy in Pakistan

The word“Literacy” has stayed alive for decades with an unchanged meaning, connotation, use, orientation and objectives. Its definition largely rests upon the teaching and learning of two basic skills: reading and writing and a brief knowledge of arithmetic. The awe-inspiring emergence and rife existence of digital tools, screen literature and digital media has significantly reshaped the concept of learning and the definition of literacy, the academicians all over the world are attempting to examine and if need be, redefine the term literacy in view of digital technology. This study was undertaken to evaluate the scope, role and influence of technology in the context of Pakistan. The data was collected from the students of English Linguistics studying at four leading Universities of Sindh including University of Sindh, Jamshoro, University of Karachi, Shah Abdul Latif University, Khairpur and Mohtarma Benazir Bhutto University, Nawabshah by applying random sampling technique. The data was processed through descriptive and inferential statistics, and the results were drawn on the basis of achieved statistics. The results reveal that students of English Linguistics in all four varsities are fast adopting digital technology mode/form of literacy. The findings show that more than 90% students use digital media. 80% students use digital tools for research and information. Only 10% students use technology for editing and proofreading. It was also learnt from the findings that despite overwhelming use of digital form of literacy by the individual youth population in Pakistan, there is however; a significant need for the academic and other professional institutions also to adopt this innovative, current and popularly proliferating form of literacy.

Research progress of diagnosis platforms based on CRISPR/Cas system

At present, nucleic acid testing technology has been widely used in clinical laboratory diagnosis. Conventional detection technique such as real-time PCR is complicated, time consuming, and dependent on specific instruments. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas) system is an adaptive immune defense system against viruses in bacteria and archaea, which has been developed into a powerful technology for genome editing. Recently, the leading groups engaged in CRISPR have set up new tools for nucleic acid detection based on Cas13a, Cas12a and newly discovered protein-Cas14, which plays an important role in rapid diagnosis of infectious diseases, detection of gene mutations in cancer and genotyping. Since they are ultrasensitive, specific, rapid and cost-effective, it is expected to bring great potential for molecular diagnosis. In this review, the mechanism of CRISPR/Cas system and the principle, the applications of the newly-developed diagnostic platforms are introduced. What′s more, the advantages, limitations and prospects of the technologies are summarized. Key words: CRISPR-Cas systems; CRISPR-associated proteins; Endonucleases; Sensitivity and specificity