Use Of Gene Editing Technologies Research Articles

Reflection on Gene Editing from the Perspective of Biopolitics

The study examines the creation of gene-edited infants from the perspective of biopolitics. Through an analysis at the level of “body-power”, we show that the infants are a product of an advanced stage of biopolitics. On the other hand, considering the level of “space-power”, we indicate that the mechanism of space deepens the governance of population through biopower, leading to real conflicts between past and future in the present. The infants can be seen as “heterotopias of mirrors”, where super-reality replaces the reality, culminating in a rational dilemma. We must also consider how to maintain our self-contemplation and naturalness when faced with the physical nature of humans and how to ensure that the state is fulfilling its role in regulating the use of gene-editing technology. Ultimately, we need to engage in a deeper rethinking and criticism of modernity to safeguard our values from being lost in the tide of modernization.

Generation and utilization of endostatin-sensitive cell lines for assessing the biological activity of endostatin.

Recombinant proteins are gaining increasing popularity for treating human diseases. The clinical effectiveness of recombinant proteins is directly related to their biological activity, which is an important indicator in drug development and quality control. However, certain recombinant proteins have unclear or complex signal pathways, making detecting their activity in vitro difficult. For instance, recombinant human endostatin (endostatin), a new antitumor drug developed in China, lacks a sensitive and stable assay for its biological activity since being market approval. To address this issue, we performed a genome-wide screening of immortalized human umbilical vein endothelial cells (HUVECs) using a CRISPR/Cas9 knockout library containing 20,000 targeted genes. We identified two potential endostatin-resistant genes, NEPSPP and UTS2, and successfully constructed a highly sensitive cell line, HUVEC-UTS2-3#, by knocking down the UTS2 gene. Based on the optimized parameters of HUVEC-UTS2-3# cells, we established a new method for detecting the biological activity of endostatin. The method was validated, and it produced results consistent with primary HUVEC cells but with higher sensitivity and more stable data. The use of gene-editing technology provides a novel solution for detecting the biological activity of recombinant proteins that other methods cannot detect.

CRISPR/Cas9-Mediated Editing of BmEcKL1 Gene Sequence Affected Silk Gland Development of Silkworms (Bombyx mori).

The silkworm (Bombyx mori) has served humankind through silk protein production. However, traditional sericulture and the silk industry have encountered considerable bottlenecks and must rely on major technological breakthroughs to keep up with the current rapid developments. The adoption of gene editing technology has nevertheless brought new hope to traditional sericulture and the silk industry. The long period and low efficiency of traditional genetic breeding methods to obtain high silk-yielding silkworm strains have hindered the development of the sericulture industry; the use of gene editing technology to specifically control the expression of genes related to silk gland development or silk protein synthesis is beneficial for obtaining silkworm strains with excellent traits. In this study, BmEcKL1 was specifically knocked out in the middle (MSGs) and posterior (PSGs) silk glands using CRISPR/Cas9 technology, and ΔBmEcKL1-MSG and ΔBmEcKL1-PSG strains with improved MSGs and PSGs and increased silk production were obtained. This work identifies and proves that BmEcKL1 directly or indirectly participates in silk gland development and silk protein synthesis, providing new perspectives for investigating silk gland development and silk protein synthesis mechanisms in silkworms, which is of great significance for selecting and breeding high silk-yielding silkworm varieties.

Application of CRISPR/Cas9 in the management of Alzheimer's disease and Parkinson's disease: a review.

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) nucleases system (CRISPR/Cas9) is a popular gene-editing technology with an expanding scope in the field of medicine. Recent studies have investigated the role of CRISPR/Cas9 system in the treatment of neurodegenerative diseases such as Alzheimer's disease (AD) and Parkinson's disease (PD). Since the risk of occurrence of both conditions is strongly associated with genetic mutations and variations, the use of gene-editing technologies to rectify these genetic errors becomes relevant. The CRISPR/Cas9 system has been tested in AD, which has led to a decrease in either amyloid beta deposition or tau phosphorylation in cells. Likewise, genetic mutations in cells affected by PD have been corrected with promising results in initial studies undertaken. Therefore, the use of the CRISPR/Cas9 system should be expanded among different populations to understand its efficacy and safety in depth among neurodegenerative conditions.

An Exploration of the Effects of Gene-Editing Technology on Human Identity

An Exploration of the Effects of Gene-Editing Technology on Human Identity

An Overview of Structured Lipid in Food Science: Synthesis Methods, Applications, and Future Prospects

Structured lipids have attracted significant interest in industries, such as food, pharmaceuticals, and cosmetics, because they can be alternatives to traditional lipids. Enzymatic synthesis techniques utilizing lipases have gained attention for structured lipids synthesis because of their site and substrate specificity. However, most lipases discovered to date exhibit sn-1,3 regiospecificity or nonspecificity, limiting the selective range of natural oils and fats that can be used as raw materials. In this review, we provide an overview of the current synthesis methods and applications of structured lipids and the limitations of existing lipases in producing structured lipids. In addition, we discuss innovative approaches, including metagenomics, and machine learning, to discover, and classify new lipases and the use of gene editing technologies for lipase engineering. These efforts aim to overcome the limitations of existing lipases and expand the range of natural oils and fats that can be used for the production of structured lipids. Therefore, this study aims to promote a better understanding of structured lipids as a material with high-added value and provides insight into ongoing research efforts in the field.

An overview of genome engineering in plants, including its scope, technologies, progress and grand challenges.

Genome editing is a useful, adaptable, and favored technique for both functional genomics and crop enhancement. Over the years, rapidly evolving genome editing technologies, including clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas), transcription activator-like effector nucleases (TALENs), and zinc finger nucleases (ZFNs), have shown broad application prospects in gene function research and improvement of critical agronomic traits in many crops. These technologies have also opened up opportunities for plant breeding. These techniques provide excellent chances for the quick modification of crops and the advancement of plant science in the future. The current review describes various genome editing techniques and how they function, particularly CRISPR/Cas9 systems, which can contribute significantly to the most accurate characterization of genomic rearrangement and plant gene functions as well as the enhancement of critical traits in field crops. To accelerate the use of gene-editing technologies for crop enhancement, the speed editing strategy of gene-family members was designed. As it permits genome editing in numerous biological systems, the CRISPR technology provides a valuable edge in this regard that particularly captures the attention of scientists.

Structural Elements of DNA and RNA Eukaryotic Expression Vectors for In Vitro and In Vivo Genome Editor Delivery.

Gene editing with programmable nucleases opens new perspectives in important practice areas, such as healthcare and agriculture. The most challenging problem for the safe and effective therapeutic use of gene editing technologies is the proper delivery and expression of gene editors in cells and tissues of different organisms. Virus-based and nonviral systems can be used for the successful delivery of gene editors. Here we have reviewed structural elements of nonviral DNA- and RNA-based expression vectors for gene editing and delivery methods in vitro and in vivo.

Structural Elements of DNA and RNA Eukaryotic Expression Vectors for In Vitro and In Vivo Genome Editor Delivery

Gene editing with programmable nucleases opens new perspectives in important practice areas, such as healthcare and agriculture. The most challenging problem for the safe and effective therapeutic use of gene editing technologies is the proper delivery and expression of gene editors in cells and tissues of different organisms. Virus-based and nonviral systems can be used for the successful delivery of gene editors. Here we have reviewed structural elements of nonviral DNA- and RNA-based expression vectors for gene editing and delivery methods in vitro and in vivo.

Comparison of public discussions of gene editing on social media between the United States and China.

The world’s first gene-edited babies event has stirred controversy on social media over the use of gene editing technology. Understanding public discussions about this controversy will provide important insights about opinions of science and facilitate informed policy decisions. This study compares public discussion topics about gene editing on Twitter and Weibo, as wel asthe evolution of these topics over four months. Latent Dirichlet allocation (LDA) was used to generate topics for 11,244 Weibo posts and 57,525 tweets from September 25, 2018, to January 25, 2019. Results showed a difference between the topics on Twitter versus Weibo: there were more nuanced discussions on Twitter, and the discussed topics between platforms focused on different areas. Temporal analysis showed that most discussions took place around gene-edited events. Based on our findings, suggestions were provided for policymakers and science communication practitioners to develop more effective communication strategies toward audiences in China and the U.S.

Gene-Edited Meat: Disentangling Consumers' Attitudes and Potential Purchase Behavior.

Novel gene-editing (GE) technologies provide promising opportunities to increase livestock productivity and to tackle several global livestock production sustainability and food security challenges. However, these technologies, as with previous genetic modification technologies in food production, are very likely to generate social controversy and opposition toward their use in the meat industry. Here, we explored public attitudes and consumption predisposition toward gene-edited meat products and their potential added benefits to livestock farming. Our results show that societal perception currently comes as a package, where the use of gene-editing technology acts as an extrinsic cue of meat products quality, and is used to make a range of inferences about all quality facets at once. Although consumers with anti-GE attitudinal positions generally were not sensitive to price discounts or added benefits, added benefits increased the consumption predisposition of most moderate and pro-GE consumers, where benefits related to animal welfare had larger effects than those relating to the environment or human health issues.

Development of antigen-receptor modified allogeneic T cell from iPS cells for cancer immunotherapy

The efficacy of T-cell therapy depends on the maintenance of antigen specificity, memory phenotype, longterm viability, and proliferative capacity of T cells in vivo. Personalized autologous T-cell therapies pose a few manufacturing challenges, in terms of quality, and supply stability. Recently, it has become possible to derive CD8 killer T cells from induced pluripotent stem cells (iPSCs) and develop CAR-CD8 killer T cells from allogeneic iPSCs. This article reviews CD8 killer T-cell induction from iPSCs, attempts to enhance process safety and reliability, and discusses the use of gene-editing technology for reducing allogeneic antigenicity.

GLUTEN FREE WHEAT (TRITICUM AESTIVUM L.): A GENETIC WAY TO CURB COELIAC DISEASE

There are three main cereal crops, rice, maize and wheat, in which, latter constitutes the major portion in the category. According to 2017 reports, it is known to be harvested around 772 million tonnes. Wheat is present in a diverse range and gets its elasticity, viscosity and properties of binding by the storage protein, Gluten, which is a secretory protein synthesised on rough endoplasmic reticulum. Gluten is roughly 78-85% of total wheat protein and made of gliadin and glutenins. There is a part of population with genes which develop autoimmune response against gluten that majorly affects the small intestine by damaging its lining and have to suffer from problems like pain in abdomen, joints, anaemia, osteoporosis and various gastro intestinal problems, developmental problems and problems like cramping, itching and lactose tolerance and excess weight loss. The patients who suffer from this disease undergo either biopsy of small intestine or serological assay like Rapid or ELISA for diagnosis. There are currently no treatments available for coeliac disease as the only way to curb this is strict gluten-free diet. There have been advances made in the field of agriculture, which resulted in production of the low gliadin content by use of gene editing technology and RNAi but the customer acceptance is the major issue faced. The most recent technological advancements in the field of biotechnology, like CRISPR-Cas9 is capable of producing the genetically modified wheat with lowest gliadin composition to curb the celiac disease impact in population.

Lysine biofortification of crops to promote sustained human health in the 21st century.

Crop biofortification is pivotal in preventing malnutrition, with lysine considered the main limiting essential amino acid (EAA) required to maintain human health. Lysine deficiency is predominant in developing countries where cereal crops are the staple food, highlighting the need for efforts aimed at enriching the staple diet through lysine biofortification. Successful modification of aspartate kinase (AK) and dihydrodipicolinate synthase (DHDPS) feedback inhibition has been used to enrich lysine in transgenic rice plants without yield penalty, while increases in the lysine content of quality protein maize have been achieved via marker-assisted selection. Here, we reviewed the lysine metabolic pathway and proposed the use of metabolic engineering targets as the preferred option for fortification of lysine in crops. Use of gene editing technologies to translate the findings and engineer lysine catabolism is thus a pioneering step forward.

Framing and legitimating EU legal regulation of human gene-editing technologies: key facets and functions of an imaginary.

ABSTRACTGene-editing technologies, ie those able to make changes in the DNA of an organism, are the object of global competition and a regulatory race between countries and regions. There is an attempt to craft legal frameworks protective enough for users, but flexible enough for developers of gene-editing technologies. This article examines the imaginary built into the framing of EU-level legal regulation of human gene-editing technologies and identifies its three key related facets: the tension around naturalness; safeguarding morality and ethics; and the pursuit of medical objectives for the protection of human health. Concerns around the use of gene-editing technologies in relation to eugenics and human enhancement have produced a multifaceted imaginary. We argue that this imaginary not only places a limit on EU-level regulation, despite a strong EU competence in respect of the internal market, but also seeks to ensure its legitimation.

Between a rock and a hard place: Farmers' perspectives on gene editing in livestock agriculture in Bavaria.

Farmers' opinions and concerns are rarely considered in public debates about the use of gene-editing technologies to modify farm animals.

Efficient CRISPR/Cas9-mediated genome editing in Rehmannia glutinosa.

Here, we cloned a phytoene desaturase (PDS) gene from Rehmannia glutinosa, and realized RgPDS1 knock out in R. glutinosa resulted in the generation of albino plants. Rehmannia glutinosa is a highly important traditional Chinese medicine (TCM) with specific pharmacologyand economic value. R. glutinosa is a tetraploid plant, to date, no report has been published on gene editing of R. glutinosa. In this study, we combined the transcriptome database of R. glutinosa and the reported phytoene desaturase (PDS) gene sequences to obtain the PDS gene of R. glutinosa. Then, the PDS gene was used as a marker gene to verify the applicability and gene editing efficiency of the CRISPR/Cas9 system in R. glutinosa. The constructed CRISPR/Cas9 system was mediated by Agrobacterium to genetically transform into R. glutinosa, and successfully regenerated fully albino and chimeric albino plants. The next-generation sequencing (NGS) confirmed that the albino phenotype was indeed caused by RgPDS gene target site editing, and it was found that base deletion was more common than insertion or replacement. Our results revealed that zCas9 has a high editing efficiency on the R. glutinosa genome. This research lays a foundation for further use of gene editing technology to study the molecular functions of genes, create excellent germplasm, accelerate domestication, and improve the yield and quality of R. glutinosa.

The Mode of Action of Endosidin20 Differs from That of Other Cellulose Biosynthesis Inhibitors.

Endosidin20 (ES20) was recently identified as a cellulose biosynthesis inhibitor (CBI) that targets the catalytic domain of CELLULOSE SYNTHASE 6 (CESA6) and thus inhibits the growth of Arabidopsis thaliana. Here, we characterized the effects of ES20 on the growth of other plant species and found that ES20 is a broad-spectrum plant growth inhibitor. We tested the inhibitory effects of previously characterized CBIs (isoxaben, indaziflam and C17) on the growth of Arabidopsis cesa6 mutants that have reduced sensitivity to ES20. We found that most of these mutants are sensitive to isoxaben, indaziflam and C17, indicating that these tested CBIs have a different mode of action than ES20. ES20 also has a synergistic inhibitory effect on plant growth when jointly applied with other CBIs, further confirming that ES20 has a different mode of action than isoxaben, indaziflam and C17. We demonstrated that plants carrying two missense mutations conferring resistance to ES20 and isoxaben can tolerate the dual inhibitory effects of these CBIs when combined. ES20 inhibits Arabidopsis growth in growth medium and in soil following direct spraying. Therefore, our results pave the way for using ES20 as a broad-spectrum herbicide, and for the use of gene-editing technologies to produce ES20-resistant crop plants.

Beyond Mendelian Genetics: Anticipatory Biomedical Ethics and Policy Implications for the Use of CRISPR Together with Gene Drive in Humans.

Clustered regularly interspaced short palindromic repeats (CRISPR) genome editing has already reinvented the direction of genetic and stem cell research. For more complex diseases it allows scientists to simultaneously create multiple genetic changes to a single cell. Technologies for correcting multiple mutations in an in vivo system are already in development. On the surface, the advent and use of gene editing technologies is a powerful tool to reduce human suffering by eradicating complex disease that has a genetic etiology. Gene drives are CRISPR mediated alterations to genes that allow them to be passed on to subsequent populations at rates that approach one hundred per cent transmission. Therefore, from an anticipatory biomedical ethics perspective, it is possible to conceive gene drive being used with CRISPR to permanently ameliorate aberrant genes from wild-type populations containing mutations. However, there are also a number of possible side effects that could develop as the result of combining gene editing and gene drive technologies in an effort to eradicate complex diseases. In this paper, we critically analyse the hypothesis that the combination of CRISPR and gene drive will have a deleterious effect on human populations from an ethical perspective by developing an anticipatory ethical analysis of the implications for the use of CRISPR together with gene drive in humans.

Shaping the CRISPR Gene-Editing Debate: Questions About Enhancement and Germline Modification.

Today's debate about the use of gene-editing technologies to alter human DNA brings together two longstanding lines of inquiry in bioethics: the ethics of human enhancement, and the ethics of heritable genetic modification. This article traces that lineage by identifying key distinctions and ethics questions in these preexisting lines of inquiry that are also employed in four recent policy and ethics statements on human gene editing. These distinctions and ethics questions can be helpful heuristics for organizing discussion, learning from existing analysis, and highlighting what is at stake with new gene-editing technologies. Yet scientists, policymakers, and others new to the ethics of emerging technologies should also be aware of both the limitations of these distinctions and past challenges in adequately addressing the ethics questions they raise. In particular, the treatment-enhancement distinction and the somatic-germline distinction are not as clear-cut as they might initially appear. More importantly, they cannot be used to definitively differentiate right from wrong uses of the technologies in question.