Genetically Modified Organisms Research Articles

Combination of upstream primer-multuplex PCR (UP-mpcr) and capillary electrophoresis for equine genetic analysis

The purity and content of DNA extracted from the sample is important during PCR analysis. In the conditions of our country, there are many cases of working on samples that do not meet the requirements for some reason. In such cases, there is a need to further test and develop other sensitive methods. The upstream-primer multiplex PCR (UP-mPCR technique) is known for its high specificity and fidelity, and has been used for detecting multiple food borne pathogens, meat species testing and detecting different genetically modified organism (GMO) insertions in a plant genome. The purpose of this experiment is to apply the UP-mPCR method on DNA samples that do not meet the quality requirements, and to test it on domestically produced diagnostics.We combined UP-mPCR with fragment analysis on DNA capillary electrophoresis genetic analyzer by applying fluorescent labelled upstream primers which were tested by amplifying 8 STRs on 23 low-quality equine gDNA samples. These samples had formerly undergone unsuccessful testing by domestic equine genotyping 15-plex kit. Single trial of UP-mPCR on the same samples showed successful amplification and detection of amplicons from 4-6 STRs, and their alleles were genotyped. Combining UP-mPCR and DNA capillary electrophoresis can be helpful in extreme situations such as having limited amounts of sample, or a shortage of multiple fluorescent dye oligonucleotides. There is no former report about the same method as combining UP-mPCR with fragment analysis.

ROLE OF BIOTECHNOLOGY IN FOOD SECURITY: A REVIEW

One of the most serious issues confronting the global food system is the wastage of approximately one-third of food at various points throughout the supply chain due to environmental and economic factors. Declines in production and food quality deterioration are concerns due to less awareness of the farming community and end users about the advanced technologies. Therefore, scientists face to develop cutting-edge technology to solve these problems and feed the bulging population to prevent starvation. Genetic engineering (GE) techniques can aid in several contexts to boost crop yields and quality. Biotechnology, genetic modification, and recombinant (r) deoxyribonucleic acid (DNA) technologies are significantly beneficial in pursuing chief progress in food production and supply. This latest literature review illustrates the recent advances in GE, their sources, current trends, and future. GE foods from animals, microbes, and crop plants have altered DNA and introduced modified genetic characteristics. Genetically modified organisms (GMOs) are vital parts of the industrial food system, and most packaged foods contain GMO ingredients that received engineering for resistance to pesticides and herbicides. Several issues raise red flags concerning GMOs, including safety, effects on the environment, and ineffective usage of pesticides. Many people are anxious about GMOs; however, most do not understand the problems.

Suitable Disinfectants with Proven Efficacy for Genetically Modified Viruses and Viral Vectors.

Viral disinfection is important for medical facilities, the food industry, and the veterinary field, especially in terms of controlling virus outbreaks. Therefore, standardized methods and activity levels are available for these areas. Usually, disinfectants used in these areas are characterized by their activity against test organisms (i.e., viruses, bacteria, and/or yeasts). This activity is usually determined using a suspension test in which the test organism is incubated with the respective disinfectant in solution to assess its bactericidal, yeasticidal, or virucidal activity. In addition, carrier methods that more closely reflect real-world applications have been developed, in which microorganisms are applied to the surface of a carrier (e.g., stainless steel frosted glass, or polyvinyl chloride (PVC)) and then dried. However, to date, no standardized methods have become available for addressing genetically modified vectors or disinfection-resistant oncolytic viruses such as the H1-parvovirus. Particularly, such non-enveloped viruses, which are highly resistant to disinfectants, are not taken into account in European standards. This article proposes a new activity claim known as "virucidal activity PLUS", summarizes the available methods for evaluating the virucidal activity of chemical disinfectants against genetically modified organisms (GMOs) using current European standards, including the activity against highly resistant parvoviridae such as the adeno-associated virus (AAV), and provides guidance on the selection of disinfectants for pharmaceutical manufacturers, laboratories, and clinical users.

Genetically Modified Organisms (GMO) Collision on Human Body: Pros and Cons

Genomic amendment is genetic procedure which possessions adjustments in genomic technology of entirely types of existing creatures. GMO is well-defined as Organisms in which genomic substantial (DNA) will be transformed in a technique which is not happen obviously via breeding or else normal reunification. Customers remain usually involved in significant concerning diet they ingest, as well as its basis beside that, stipulation handled, the materials that might adding towards. By means of new ideas arise in nourishment creation schemes, customers could remain further apprehensive around then concerned of buying freshly technologically advanced foodstuffs. Hereditarily planned or hereditarily improved diets, or else individuals that comprise about hereditarily improved creatures, were presented in US marketplace as well as acted proceeding superstore tables popularin 1994 by Flavr Savr tomato. FDA agreed to the technique through Calgene of introducing genetic material which avoids accumulation of an enzyme which will then reason of unstiffening in the fruitlet, letting them economically vended Flavr Savr tomato to have extended service lifetime along with conservative tomatoes . On the other hand, the aforementioned manufactureterminated in 1997. Later two years, the overview of Flavr Savr, during 1996, weed killer resistingsoybeans have being announced within foodstuff scheme via let agriculturalists towards procedure which is generally appropriate for weed killer Roundup surrounded by meadow directed towards destroy a widespread variety of tidies deprived of damaging their unaffected Roundup Ready Harvests. During that time, GM and GMOs foodstuffs during specific increase incessant community discussion by means of favors their security, hazards, regulation, tagging and limit.

A robust yeast biocontainment system with two-layered regulation switch dependent on unnatural amino acid

Synthetic auxotrophy in which cell viability depends on the presence of an unnatural amino acid (unAA) provides a powerful strategy to restrict unwanted propagation of genetically modified organisms (GMOs) in open environments and potentially prevent industrial espionage. Here, we describe a generic approach for robust biocontainment of budding yeast dependent on unAA. By understanding escape mechanisms, we specifically optimize our strategies by introducing designed “immunity” to the generation of amber-suppressor tRNAs and developing the transcriptional- and translational-based biocontainment switch. We further develop a fitness-oriented screening method to easily obtain multiplex safeguard strains that exhibit robust growth and undetectable escape frequency (<~10−9) on solid media for 14 days. Finally, we show that employing our multiplex safeguard system could restrict the proliferation of strains of interest in a real fermentation scenario, highlighting the great potential of our yeast biocontainment strategy to protect the industrial proprietary strains.

Validation and comparison of four DNA extraction methods for genetically modified organisms analysis: From DNA quality control to quantification of genetically modified content in real‐life samples matrices

AbstractBackgroundThe analysis of genetically modified organisms (GMOs) in food and feed is broadly based on the detection of target DNA sequences using PCR‐based methods. The first step of a typical workflow in a GMO testing laboratory is DNA extraction and purification. Several DNA extraction methods have been described for this purpose so far. Whatever the method adopted, the yield and the quality of the extracted DNA are essential factors for the success of GMO analysis.ResultsThe DNA extraction yield was variable between the different methods, while quality and efficiency appeared comparable. Each method allowed a different yield and a slight correlation between inhibition and the food/feed matrix extracted was observed. Particularly for challenging matrices, the method used for DNA extraction has a remarkable influence on both the quality and quantity of the recovered DNA. Small variations in value % GM content were observed in complex, heterogeneous matrices but not in CRMs and homogeneous matrices.ConclusionsIn this study, the purpose is not to establish the best method of extraction but to examine several parameters that play an important role in GMO detection and verify a possible effect on the quantified GMO percentage. In particular, it was found that the yield and quality of extracted DNA depends on the type of sample, notably the degree of processing, composition, particle size, and the protocol used, especially in relation to sample intake. The slightly different GM content observed in heterogeneous matrices could raise doubt in the case of samples close to the legal threshold.

Green crimes: the impact of genetically modified organisms on promoting food security in Kenya

Kenya’s food insecurity hinders progress towards sustainable development. While the Kenyan Constitution guarantees every person the right to adequate food of acceptable quality, it also prohibits environmental and health endangerment. Whether and how to invest in genetically modified organism (GMO) technology as an alternative food production method is important. In this context, scientists should not be denied the opportunity of harmonising the tension between environmental safety and food security while upholding sustainable development. Scientifically, GMO crops are sustainable, notwithstanding the African Union’s rigid social and political setting. However, each state’s role in providing sufficient resources and law enforcement personnel is crucial. A GMO regulatory system addresses environmental safety and human health, explicitly adopting thedevelopmental risk notion. Kenya’s 2012 cabinet ban on GMO foods derived from the Séralini Report which erroneously claimed that GM maize causes cancer in rodents. The health ministry established a Task Force to review the country’s readiness regarding GMO safety and adoption. Despite having been completed in 2014, its Report remains secret. In 2015, the High Courtdismissed as premature demands for public participation on whether to unban GMOs. In 2022, President Ruto lifted the ban. Arguably, while permitting GMO experimentation, it is prudent to prescribe criminal sanctions. Beyond anthropocentric notions, green criminology provides a framework to analyse both illegal and legal environmental harms, and for appraising Kenya’s evolving GMO policy. The Constitution provides a right to sustainable use and also establishes enforcement mechanisms to compel cessation and restoration. Yet without punitive consequences, GMO regulations may not deter offenders from environmental contamination.

Detection of commercialized plant products derived from new genomic techniques (NGT) - Practical examples and current perspectives

New genomic techniques (NGT) are increasingly applied in plant research and development in many countries due to their simplicity in introducing genetic modifications that alter specific traits. First NGT products are on the market in several countries where no specific regulatory approval is required. In the European Union (EU), however, all NGT derived products are subject to the existing genetically modified organism (GMO) authorization regulations. This nourishes the discussion on the technical and analytical feasibility of detecting, identifying and differentiating plants produced by targeted mutagenesis from conventional breeding products and spontaneous mutations in routine food/feed and seed/grain testing by enforcement laboratories. The anticipated difficulties are currently based mainly on theoretical considerations, particularly with regard to single nucleotide or very limited sequence changes. Against that background, this brief review examines concrete and practical approaches for the detection and identification of market-relevant NGT plants. Four early commercialized NGT plants have been examined as real examples to verify the feasibility of detection method design and development using routinely applied analytical approaches. In addition, we searched for potential new screening approaches and show, how a common and well conserved genetic element used in CRISPR/Cas9 applications may serve as a potential target, if this foreign DNA element is still integrated in the host genome. The conclusions to be drawn from the considered theoretical approaches and the practical examples are described in the context of key differences to the current GMO testing practice, in particular regarding the difficulty to develop methods specifically identifying NGT applications. Finally, we discuss whether the consideration of such factors could facilitate a resolution of the current dilemma between detection and the ability to identify plant NGT events.

Belgian dietitians' knowledge, perceptions and willingness-to-recommend of genetically modified food and organisms.

Dietitians play a critical role in the public's relationship with food and are often overlooked as an important stakeholder group in the general debate about sustainable food. Genetically modified organisms (GMOs) are one type of modern food source that could contribute to a more sustainable food system. This case study is the first to examine the knowledge, perception and willingness-to-recommend (WTR) genetically modified (GM) foods by dietitians in Europe. An online survey was addressed to all members of the Flemish Association of Dietitians (Belgium) in 2021, resulting in a sample of 98 valid responses. Multivariate linear regression included sociodemographic, knowledge, and attitudinal factors as the independent variables to explain dietitians' WTR. Flemish dietitians had limited knowledge of GMOs; only about half of the GM questions were answered correctly. Most dietitians (53%-76%) would recommend GMOs with positive effects on human nutrition or sustainability, whereas few dietitians (19%-27%) would recommend other GMO applications. Trust in GMO information sources and perceived GM benefits significantly influenced a positive WTR of GM foods. Predominant negative information about GM foods was significantly associated with dietitians' low trust and WTR such foods. Countering the predominantly negative portrayal with more neutral and factual information could improve trust, which in turn could positively influence dietitians' perceptions towards GMOs. By further examining the knowledge and perception of dietitians worldwide GMOs and gene-edited products, new insights could be could gathered into the positioning of this underexposed stakeholder group.

Critical Reading of Informational Texts (CRIT) Scaffold: Evaluating the efficacy of an instructional scaffold for reading multiple scientific texts

Developing critical processing of online information is one important mission of schooling in the 21st century. However, science teachers often lack an instructional tool they can integrate into existing curricula to support students’ information processing. This study aims to examine the efficacy of the Critical Reading of Informational Texts (CRIT) scaffold on students’ critical integrative argumentation - the dialogic process of weighing, evaluating, and integrating scientific claims. The present study documented the design of the CRIT scaffold to support both critical evaluation and integration of online scientific information. Initial efficacy evidence from a cluster-randomized control group study demonstrated that students who used the scaffold produced written task products that had higher overall argumentative quality and were more likely to support evidence-based conclusions about Genetically Modified Organisms (GMOs), weigh claims using source and evidence quality, and engage in refutation by countering specific claims based on why they are flawed.

Reproducibility of next-generation-sequencing-based analysis of a CRISPR/Cas9 genome edited oil seed rape

Next-generation-sequencing (NGS) becomes increasingly important for laboratories tasked with the detection of genetically modified organisms (GMOs) in food, feed and seeds. Its implementation into standardized workflows demands reliable intra- and inter-laboratory reproducibility. Here, we analyze the reproducibility of short- and long-read targeted NGS and long-read whole genome sequencing (WGS) data between three independent laboratories. Replicate samples were submitted for sequencing and comparatively analyzed. The targeted-NGS-samples consisted of oil seed rape (OSR) sampled from a commodity shipment spiked with a genome edited (GE) OSR and the WGS-samples consisted of leaf material from the GMOs’ parental line. All laboratories delivered highly reproducible high-quality targeted NGS data with little variation. The detection of GMO-related sequences works well regardless of the facility, while the mapping to the complex genome is superior using long read data. Long read WGS is currently not suitable for routine use in enforcement laboratories, due to a large inter-laboratory variation.

Beyond plastic – Consumers prefer food packaging derived from genetically modified plants

The technology of genetically modified organisms (GMO) and especially genetically modified plants (GMP) applied in agriculture is a key element in a bioeconomy-based sustainability transition. However, consumers in many European countries allegedly disapprove of ‘green biotechnology’. Drawing on innovation adoption and risk perception literature, we explore the acceptance of GMP-based food packaging. Fossil-fuel-based production and plastic waste have become a major concern in many societies, and GMP-based technologies might offer solutions. Taking fully degradable material based on GMP as an example to replace conventional plastic in many applications, we present new insights into the GMP debate using a choice-based conjoint analysis in four European countries. Our results show that GMP-based packaging material is preferred to conventional plastic packaging in all countries. We present implications for research policies.

Assessing the environmental fate of rAAV in activated sludge and water: Implications for environmental risk assessments and GMO regulatory frameworks

During the past +20 years, recombinant adeno-associated virus (rAAV) vectors have emerged as the primary vehicle of choice for in vivo gene therapy. rAAV vectors are classified as genetically modified organisms (GMOs), therefore specific biosafety laws apply regarding their use. Environmental agencies participating in the review of clinical trial applications involving viral-based gene therapies (eg based on AAV) focus among other phenomena especially on shedding, a mechanism by which rAAV vectors exit a patient's body and enter the natural environment. For example, following patient use, shed viral particles excreted in urine and feces enter the wastewater treatment facility (WWTF) and subsequently may be released into the natural environment through wastewater effluent discharges. Based on basic molecular biology, it is generally assumed by the scientific community that shed rAAV particles will undergo degradation during the wastewater treatment process. However, despite their importance and increase in use during the last few decades, actual data to support our understanding of the environmental fate of shed rAAV vector particles is unavailable. Data to support this assumption would greatly enhance our knowledge and understanding of degradation kinetics associated with rAAV in the environment. Such data would also provide strong scientific support for changes in current legislation regarding the medicinal use of GMOs. Therefore, the goal of this research was to conduct laboratory experiments to assess the actual environmental fate of rAAV virions.In this study the stability of 4 different rAAV vectors (based on wildtype (wt) AAV serotypes 2,3,6,9) was assessed during incubation in activated sludge (containing live microorganisms). This setting corresponds to conditions as encountered in WWTFs, and has been used in order to assess rAAV fate under environmentally relevant conditions, to gain a better understanding of the general environmental risk posed by shed rAAV particles. The amount of detectable virions in the supernatant, as measured by sensitive and specific qPCR, rapidly decreased within hours and continued to decline, reaching the lower limit of quantitation prior to or by study termination on day 7. Furthermore, a half-life of approximately 7 days for rAAV virions was determined under abiotic conditions, during a room temperature incubation experiment of rAAV vectors in water in the absence of any microbiota or sludge.The findings from this study provide the first insight of its kind into the actual environmental fate of shed rAAV particles, and help the community to better understand the potential impact of rAAVs on the environment. It has become evident now that shed particles are not equipped to remain stable and/or soluble once entering a typical WWTF and therefore do not pose a threat to the natural environment. These findings support a data-driven approach towards a simplified, risk-based regulation of medicinal GMOs in the EU and other regions.

Public Discourse of Genetically Modified Organisms in China: An Investigation of Commenting and Reposting Behaviors on Social Media

This study examined the factors affecting user engagement in the genetically modified organism (GMO) discourse on one of the most popular social media platforms in China. A content-analysis study was conducted with a sample of the most commented GMO posts on social media and over 7000 comments. Firstly, posts from well-known and government accounts facilitated the online discourse of GMOs. Secondly, the nature of the event and the features of GMO messages influenced how users engaged with the online discussion. Thirdly, among the commenters, female users were more likely to agree with the original posts whereas male users were more likely to repost the message. In conclusion, social media users held negative attitudes toward GMOs as they tended to repost more and agree more when GMO risks were mentioned. Source and message features, as well as commenters’ characteristics, were found to have a significant impact on user engagement. Social media continued to serve as an important public space to facilitate public debate on a controversial scientific topic.

Agricultural terrorism and food insecurity in Indonesia: a policy analysis

As a form of bioterrorism, agricultural terrorism/agroterrorism leads to livestock and crop interventions. Agroterrorism is part of economic warfare, which involves the destruction of a country's agricultural or ecological infrastructure using biological weapons. Agroterrorism leads to a country's food insecurities and the collapse of its economic climate. The development of biotechnology allows the modification of biological elements to result in changes in the characteristics of living things infiltrated by these agents. Genetically Modified Organism (GMO) technology, for instance, is a paradoxical double-edged sword. GMOs can generate economic benefits, but can also result in huge economic losses if they are weaponized as agroterrorism. Using a qualitative approach, this study conducts a policy analysis to explain the factors that impact the absence of food safety policies from agroterrorism threats in Indonesia. Data collection was conducted through literature searches in the form of articles, documents, and policies with the keywords "agricultural terrorism"; "Genetically Modified Organism"; "food insecurity"; "biotechnology", and "national security"; as well as by analyzing the experiences of other countries. As a country with direct land borders with other countries, Indonesia needs extra efforts to secure the country's biodiversity. As one of the consequences of the ASEAN-China free trade, the possibility of agroterrorism events has become more likely to occur in Indonesia. The development of a solid food infrastructure may overcome this problem. Recommendations for safe and resilient food policy scenarios should be pursued to ensure quality food production and sustainable economic growth for a better Indonesia, not only in the present, but for the sake of sustainability in the future.

Central–Local Relations in China: A Case Study of Heilongjiang's GMO Ban

AbstractThis study contributes to the research on central–local relations in China by examining local dynamics and defiance. Drawing on the case of a provincial government's defiance against a central policy – Heilongjiang province's 2016 ban on genetically modified organisms (GMOs) – this study shows that despite the unprecedented recentralization push in recent years, local defiance still exists and persists. In addition, this study finds that the Heilongjiang provincial government managed to reduce potential political backlash by feeding the public distrust of GMOs, exploiting the internal divide and central ambiguity over GMOs and, more importantly, skilfully framing its GMO ban as part of its efforts to implement Xi Jinping's Green Development Concept.

GMO discussion on Twitter

ABSTRACT This paper focuses on analyzing discussions related to Genetically Modified Organisms (GMOs) on Twitter, with a specific focus on the spread of misinformation and conspiracy theories. The authors collected and analyzed 1,048,274 English tweets related to GMOs between January 2020 and December 2022 using the Twitter API. The tweets were subjected to topical and sentiment analysis to identify the prevalent themes and attitudes toward GMOs. 30.92% of the tweets in the observed period were negative, 21.65% were neutral, and 47.43% were positive. The authors identified four clusters of tweets associated with misinformation or conspiracy theories: GMOs and vaccines, GMOs and COVID-19, GMOs and Monsanto, and GMOs and Bill Gates. The findings of this analysis can inform strategies for combating the spread of false information and conspiracies on social media and improve public understanding and trust in GMO technology.

Rapid Visual LAMP Method for Detection of Genetically Modified Organisms.

We developed a novel loop-mediated isothermal amplification (LAMP) method using DNA captured on polyacrylamide microparticles (PAMMPs) as templates (PAMMPs@DNA-LAMP) for rapid qualitative detection of genetically modified organisms (GMOs). Here, DNA was extracted by a fast and cost-effective method using PAMMPs. Four LAMP primers were designed for the PAMMPs@DNA-LAMP method to detect the cauliflower mosaic virus 35S (CaMV35S) promotor in GMOs. We thus developed this method for rapid extraction of DNA (5-10 min) and fast amplification of DNA within ∼30 min at a constant temperature of 63 °C. Moreover, the DNA captured by PAMMPs (PAMMPs@DNA) could be effectively detected by both conventional and quantitative PCR (qPCR) and LAMP. The PAMMPs@DNA-LAMP method was validated with high specificity, sensitivity, and performance for practical sample analysis. This assay detected 0.01% target sequences, which had a high specificity like qPCR and better than the conventional PCR (cPCR). Furthermore, PAMMPs@DNA-LAMP was successfully used to extract and detect DNA from food samples of the major crops (soybean, maize, rice, etc.). In summary, a novel PAMMPs@DNA-LAMP assay has been developed, which has higher sensitivity and spends less time than the cPCR detection using the conventional DNA extracted process. This method offers a novel approach for rapid detection of GMOs in the field.

Towards the Phenomenology of Hybrids as Regenerative Design and use – a Post-Heideggerian Account

Grasping the identity of hybrids, that is beings which cross the binarism of nature and technology (e.g. genetically-modified organisms (GMOs), synbio inventions, biomimetic projects), is problematic since it is still guided by self-evident dualistic categories, either as artefacts or as natural entities. To move beyond the limitations of such a one-sided understanding of hybrids, we suggest turning towards the categories of affordances and the juxtaposition of needs and patterns of proper use, as inspired by the Heideggerian version of phenomenology. Drawing upon selected concepts by Heidegger, we argue that hybrids can be conceptualised as a regenerative design and use to serve the planet. We argue that the ideal type of non-exploitative account of hybrids consists of the adaptive approach to the environment, which does not, however, exclude the possibility of designing and constructing new beings. We also point out that hybrids undermine the divide of being destructive/regenerative which marks the boundaries of nature and technology.

Establishment of monoclonal antibody and scFv immuno-based assay for Cry2Aa toxin in spiked grain samples

Bacillus thuringiensis (Bt) Cry toxins have been widely used in the development of genetically modified organisms (GMOs) for pest control. This work aimed to establish more cost effective methods for used Cry2Aa toxins. Three immunoassay methods (IC-ELISA, DAS-ELISA, and CLEIA) were successfully developed in this work. The mAb was used as the detecting antibody, for the IC-ELISA, the range of IC20 to IC80 was 1.11 μg/mL - 60.70 μg/mL, and an IC50 of 10.65 μg/mL. For the DAS-ELISA, the limit of detection (LOD) and limit of quantitation (LOQ) were 10.76 ng/mL and 20.70 ng/mL, respectively. For the CLEIA, the LOD and LOQ were 6.17 ng/mL and 7.40 ng/mL, respectively. The scFv-based detections were the most sensitive for detecting Cry2Aa. The LOD and LOQ for the DAS-ELISA were 118.75 ng/mL and 633.48 ng/mL, respectively. The LOD and LOQ for the CLEIA, read as 37.47 ng/mL and 70.23 ng/mL, respectively. The fact that Cry2Aa toxin was recovered in spiked grain samples further demonstrated that the approaches might be used to identify field samples. These methods provided good sensitivity, stability, and applicability for detecting Cry2Aa toxin, promising ultrasensitive monitoring and references for Cry toxins risk assessment.