scholarly journals Production and Purification of Artificial Circular RNA Sponges for Application in Molecular Biology and Medicine

2020 ◽  
Vol 3 (2) ◽  
pp. 42 ◽  
Author(s):  
Janina Breuer ◽  
Oliver Rossbach
Keyword(s):  
Molecular Biology ◽  
Large Scale ◽  
Circular Rna ◽  
In Vitro Transcription ◽  
Circular Rnas ◽  
Small Scale ◽  
Limiting Factor ◽  
Scale Production ◽  
Naturally Occurring

Characterized by their covalently closed structure and thus an elevated stability compared to linear RNA molecules, circular RNAs (circRNAs) form a novel class of mainly non-coding RNAs. Although the biological functions of naturally occurring circRNAs are largely unknown, they were reported to act as molecular sponges, sequestering microRNAs (miRNAs), resulting in a de-repression of target mRNAs. Taking these characteristics of naturally occurring circRNAs into account, artificial circRNAs could be a potential tool in molecular biology and medicine. Using the Hepatitis C virus (HCV) as a model system, this application of artificial circular RNAs was demonstrated. The virus requires cellular miRNA miR-122 for its life cycle, and circRNAs specifically engineered to efficiently sequester this miRNA impacted viral propagation. Since in this context the production of engineered circRNA remains the limiting factor, we present a method to produce and efficiently purify artificial circRNA sponges (ciRS) in vitro. In this protocol we provide insights into a small-scale and large-scale production technique of artificial circular RNA sponges relying on in vitro transcription and RNA ligation.

scholarly journals Regeneration Technique of Bamboo Species through Nodal Segments: A Review

2020 ◽  
Vol 8 (1) ◽  
pp. 54-68
Author(s):  
Meena Maiya Suwal ◽  
Janardan Lamichhane ◽  
Dhurva Prasad Gauchan
Keyword(s):  
Large Scale ◽  
Physiological Stress ◽  
Perennial Grass ◽  
Shoot Multiplication ◽  
Nodal Segments ◽  
Individual Species ◽  
Small Scale ◽  
Scale Production ◽  
Bamboo Species

Micropropagation is an alternative technique to propagate at large scale plants to meet global plant demand. Various researchers have worked on the micropropagation technique to regenerate bamboo species by using nodal segments from years. Contamination, browning, necrosis, and acclimatization with physiological stress are the extreme problems of the micropropagation technique. But, many numbers of papers have been published on micropropagation of the bamboo species through nodal segments as explants. The proliferation of the bamboo shoots is dependent on the season of collection, size of explants, the position of explants, diversity of plants, concentration and combination of plant growth regulators, most adequate culture medium, environmental condition of the equipment, handling, and individual species. Bamboo is a monocarpic fast-growing, tall perennial grass and having the high potential to generate economic and social benefits. It helps to maintain land patterns and control soil erosion.  The long life cycle of the bamboo produces a huge amount of seeds but unfortunately, mostly, they are non-viable. So, bamboos are propagated from vegetative by cutting and air layering. However, these methods are only for a small scale and they also tend to destroy large mother plant stocks and difficult to be transported. So, the in vitro propagation technique is useful to obtain large progenies from desired genotypes. Mostly, BAP and TDZ growth hormones are widely used for shoot multiplication and IBA, NAA and IAA are used for root initiation as per developed protocols in tissue culture for large scale production. This review intends to explore an overview of the recent literature reports to summarize the importance of micropropagation by using nodal segments of bamboo species and factors influencing it.

scholarly journals Large-scale circular RNA deregulation in T-ALL: unlocking unique ectopic expression of molecular subtypes

2020 ◽  
Vol 4 (23) ◽  
pp. 5902-5914
Author(s):  
Alessia Buratin ◽  
Maddalena Paganin ◽  
Enrico Gaffo ◽  
Anna Dal Molin ◽  
Juliette Roels ◽  
...  
Keyword(s):  
Large Scale ◽  
Molecular Subtypes ◽  
Lymphoblastic Leukemia ◽  
Molecular Genetic ◽  
Ectopic Expression ◽  
Circular Rna ◽  
Circular Rnas ◽  
Sequencing Data ◽  
Significant Differential Expression

Abstract Circular RNAs (circRNAs) are stable RNA molecules that can drive cancer through interactions with microRNAs and proteins and by the expression of circRNA encoded peptides. The aim of the study was to define the circRNA landscape and potential impact in T-cell acute lymphoblastic leukemia (T-ALL). Analysis by CirComPara of RNA-sequencing data from 25 T-ALL patients, immature, HOXA overexpressing, TLX1, TLX3, TAL1, or LMO2 rearranged, and from thymocyte populations of human healthy donors disclosed 68 554 circRNAs. Study of the top 3447 highly expressed circRNAs identified 944 circRNAs with significant differential expression between malignant T cells and normal counterparts, with most circRNAs displaying increased expression in T-ALL. Next, we defined subtype-specific circRNA signatures in molecular genetic subgroups of human T-ALL. In particular, circZNF609, circPSEN1, circKPNA5, and circCEP70 were upregulated in immature, circTASP1, circZBTB44, and circBACH1 in TLX3, circHACD1, and circSTAM in HOXA, circCAMSAP1 in TLX1, and circCASC15 in TAL-LMO. Backsplice sequences of 14 circRNAs ectopically expressed in T-ALL were confirmed, and overexpression of circRNAs in T-ALL with specific oncogenic lesions was substantiated by quantification in a panel of 13 human cell lines. An oncogenic role of circZNF609 in T-ALL was indicated by decreased cell viability upon silencing in vitro. Furthermore, functional predictions identified circRNA-microRNA gene axes informing modes of circRNA impact in molecular subtypes of human T-ALL.

scholarly journals Mildred Rebstock: Profile of the Medicinal Chemist Who Synthesized Chloramphenicol

2019 ◽  
Vol 63 (6) ◽  
Author(s):  
David M. Aronoff
Keyword(s):  
United States ◽  
Large Scale ◽  
The United States ◽  
70Th Anniversary ◽  
Scale Production ◽  
Naturally Occurring ◽  
Large Scale Production

ABSTRACTThis year marks the 70th anniversary since Parke-Davis and Company announced the synthesis of chloramphenicol, the first naturally occurring antibiotic to be chemically generatedin vitrofor large-scale production. The effort was led by the chemist Mildred Rebstock, Ph.D., (1919 to 2011), who would turn 100 years old this year. Her accomplishment, at a time when very few chemists in the United States were women, was celebrated internationally. This commentary reviews her important contribution.

AGRICULTURAL PRODUCTION OF CA DONG PEOPLE IN RESETTLEMENT AREA OF TRANH RIVER HYDROPOWER NUMBER 2: CURRENT SITUATION AND IMPACT FACTORS

2020 ◽  
Vol 9 (2) ◽  
Author(s):  
Bùi Thị Bích Lan
Keyword(s):  
Social Relations ◽  
Agricultural Production ◽  
Large Scale ◽  
Transition Process ◽  
Impact Factors ◽  
Small Scale ◽  
Scale Production ◽  
Technical Application ◽  
Production Models ◽  
Role Of The State

In Vietnam, the construction of hydropower projects has contributed significantly in the cause of industrialization and modernization of the country. The place where hydropower projects are built is mostly inhabited by ethnic minorities - communities that rely primarily on land, a very important source of livelihood security. In the context of the lack of common productive land in resettlement areas, the orientation for agricultural production is to promote indigenous knowledge combined with increasing scientific and technical application; shifting from small-scale production practices to large-scale commodity production. However, the research results of this article show that many obstacles in the transition process are being posed such as limitations on natural resources, traditional production thinking or the suitability and effectiveness of scientific - technical application models. When agricultural production does not ensure food security, a number of implications for people’s lives are increasingly evident, such as poverty, preserving cultural identity, social relations and resource protection. Since then, it has set the role of the State in researching and building appropriate agricultural production models to exploit local strengths and ensure sustainability.

scholarly journals Lentiviral Vectors for Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1288
Author(s):  
Wendy Dong ◽  
Boris Kantor
Keyword(s):  
Large Scale ◽  
Lentiviral Vector ◽  
Clinical Applications ◽  
Scale Production ◽  
Base Editing ◽  
Epigenome Editing ◽  
Vector Systems ◽  
Dividing Cells

CRISPR/Cas technology has revolutionized the fields of the genome- and epigenome-editing by supplying unparalleled control over genomic sequences and expression. Lentiviral vector (LV) systems are one of the main delivery vehicles for the CRISPR/Cas systems due to (i) its ability to carry bulky and complex transgenes and (ii) sustain robust and long-term expression in a broad range of dividing and non-dividing cells in vitro and in vivo. It is thus reasonable that substantial effort has been allocated towards the development of the improved and optimized LV systems for effective and accurate gene-to-cell transfer of CRISPR/Cas tools. The main effort on that end has been put towards the improvement and optimization of the vector’s expression, development of integrase-deficient lentiviral vector (IDLV), aiming to minimize the risk of oncogenicity, toxicity, and pathogenicity, and enhancing manufacturing protocols for clinical applications required large-scale production. In this review, we will devote attention to (i) the basic biology of lentiviruses, and (ii) recent advances in the development of safer and more efficient CRISPR/Cas vector systems towards their use in preclinical and clinical applications. In addition, we will discuss in detail the recent progress in the repurposing of CRISPR/Cas systems related to base-editing and prime-editing applications.

scholarly journals Fabrication Techniques for Graphene Oxide-Based Molecular Separation Membranes: Towards Industrial Application

Nanomaterials ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 757
Author(s):  
Ohchan Kwon ◽  
Yunkyu Choi ◽  
Eunji Choi ◽  
Minsu Kim ◽  
Yun Chul Woo ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Large Scale ◽  
Spray Coating ◽  
Dip Coating ◽  
Small Scale ◽  
Scale Production ◽  
Separation Membranes ◽  
Current Review ◽  
Slot Die ◽  
Continuous Fabrication

Graphene oxide (GO) has been a prized material for fabricating separation membranes due to its immense potential and unique chemistry. Despite the academic focus on GO, the adoption of GO membranes in industry remains elusive. One of the challenges at hand for commercializing GO membranes lies with large-scale production techniques. Fortunately, emerging studies have acknowledged this issue, where many have aimed to deliver insights into scalable approaches showing potential to be employed in the commercial domain. The current review highlights eight physical methods for GO membrane fabrication. Based on batch-unit or continuous fabrication, we have further classified the techniques into five small-scale (vacuum filtration, pressure-assisted filtration, spin coating, dip coating, drop-casting) and three large-scale (spray coating, bar/doctor blade coating, slot die coating) approaches. The continuous nature of the large-scale approach implies that the GO membranes prepared by this method are less restricted by the equipment’s dimensions but rather the availability of the material, whereas membranes yielded by small-scale methods are predominately limited by the size of the fabrication device. The current review aims to serve as an initial reference to provide a technical overview of preparing GO membranes. We further aim to shift the focus of the audience towards scalable processes and their prospect, which will facilitate the commercialization of GO membranes.

Abstract 262: Derivation of Duchenne Muscular Dystrophy Disease Specific Cardiomyocytes From Patient Urine

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Xuan Guan ◽  
David L Mack ◽  
Claudia M Moreno ◽  
Fernando Santana ◽  
Charles E Murry ◽  
...  
Keyword(s):  
Stem Cells ◽  
Human Urine ◽  
Large Scale ◽  
Lentiviral Vector ◽  
Cellular Reprogramming ◽  
Urine Samples ◽  
Pcr Analysis ◽  
Scale Production ◽  
Mechanism Study

Introduction: Human somatic cells can be reprogrammed into primitive stem cells, termed induced pluripotent stem cells (iPSCs). These iPSCs can be extensively expanded in vitro and differentiated into multiple functional cell types, enabling faithful preservation of individual’s genotype and large scale production of disease targeted cellular components. These unique cellular reagents thus hold tremendous potential in disease mechanism study, drugs screening and cell replacement therapy. Due to the genetic mutation of the protein dystrophin, many DMD patients develop fatal cardiomyopathy with no effective treatment. The underlying pathogenesis has not been fully elucidated. Hypothesis: We tested the hypothesis that iPSCs could be generated from DMD patients’ urine samples and differentiated into cardiomyocytes, recapitulating the dystrophic phenotype. Methods: iPSCs generation was achieved by introducing a lentiviral vector expressing Oct4, Sox2, c-Myc and Klf4 into cells derived from patient’s (n=1) and healthy volunteers’ (n=3) urine. Cardiomyocytes were derived by sequentially treating iPSCs with GSK3 inhibitor CHIR99021 and Wnt inhibitor IWP4. Differentiated cardiomyocytes were subjected to calcium imaging, electrophysiology recording, Polymerase Chain Reaction (PCR) analysis, and immunostaining. Results: iPSCs were efficiently generated from human urine samples and further forced to differentiate into contracting cardiomyocytes. PCR analysis and immunostaining confirmed the expression of a panel of cardiac markers. Both normal and patient iPSC derived cardiomyocytes exhibited spontaneous and field stimulated calcium transients (up to 2Hz), as well as action potentials with ventricular-like and nodal-like characteristics. Anti-dystrophin antibodies stained normal iPSC-derived cardiomyocyte membranes but did not react against DMD iPSC-derived cardiomyocytes. Conclusions: Cardiomyocytes can be efficiently generated from human urine, through the cellular reprogramming technology. DMD cardiomyocytes retained the patient’s genetic information and manifested a dystrophin-null phenotype. Functional assessments are underway to determine differences that may exist between genotypes.

scholarly journals Thymosin-β4 Mediates Hepatic Stellate Cell Activation by Interfering with CircRNA-0067835/miR-155/FoxO3 Signaling Pathway

2018 ◽  
Vol 51 (3) ◽  
pp. 1389-1398 ◽  
Author(s):  
Lili Zhu ◽  
Tingting Ren ◽  
Zixin Zhu ◽  
Mingliang  Cheng ◽  
Qiuju Mou ◽  
...  
Keyword(s):  
Liver Fibrosis ◽  
Cell Activation ◽  
Stellate Cell ◽  
Circular Rna ◽  
Fine Tuning ◽  
Differentially Expressed ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
G1 Arrest

Background/Aims: Hepatic stellate cells (HSCs) are the primary cell type responsible for liver fibrosis. Our study proved that thymosin beta 4 (Tβ4) has anti-fibrogenic effects in HSCs through PI3K/AKT pathway. However, the underlying mechanisms are not fully elucidated. Circular RNAs (circRNAs) play important roles in fine-tuning gene expression and are often deregulated in cancers. However, the expression profile and clinical significance of in liver fibrosis is still unknown. Therefore, we hypothesize that Tβ4 influences circRNAs in liver fibrosis. Methods: Circular RNA microarray was conducted to identify Tβ4-related circRNAs. Pathway analysis and miRNA response elements analysis was conducted to predict the potential roles of differentially expressed circRNAs in liver fibrosis. CCK8 assays and flow cytometric assays were conducted to clarify the role of circRNA in liver fibrosis. Bioinformatics analysis and in vitro experiments were conducted to clarify the mechanism of circRNA-mediated gene regulation in liver fibrosis. Results: A total of 644 differentially expressed circRNAs were identified between the Tβ4-depleted LX-2 cells and the control LX2 cells. The expression of circRNA-0067835 was significantly increased in the Tβ4-depleted LX-2 cells compared with control. Knockdown of circRNA-0067835 observably decreased LX-2 cell proliferation by causing G1 arrest and promoting apoptosis. Bioinformatics online programs predicted that circRNA-0067835 acted as miR-155 sponge to regulate FOXO3a expression, which was validated using luciferase reporter assay. Conclusion: Our experiments showed that circRNA-0067835 regulated liver fibrosis progression by acting as a sponge of miR-155 to promote FOXO3a expression, indicating that circRNA-0067835 may serve as a potential therapeutic target for patients with liver fibrosis.

scholarly journals Influência da vernalização de semente na produção, crescimento e desenvolvimento de plantas de lisianthus

2018 ◽  
Vol 39 (6) ◽  
pp. 2325 ◽  
Author(s):  
Maria Yumbla-Orbes ◽  
José Geraldo Barbosa ◽  
Wagner Campos Otoni ◽  
Marcel Santos Montezano ◽  
José Antônio Saraiva Grossi ◽  
...  
Keyword(s):  
Large Scale ◽  
Limiting Factor ◽  
Scale Production ◽  
Cut Flower ◽  
Cut Flowers ◽  
Flower Production ◽  
Commercial Scale ◽  
Large Scale Production ◽  
And Control

Flowering induction and control is a limiting factor when commercially producing cut flowers of lisianthus and seed exposure to low temperatures, a physiological event called vernalization, induces the differentiation of vegetative buds to reproductive buds, contributing to a flowering that is uniform and has quality. The objective of this study was to evaluate the influence of seed vernalization in three cultivars of lisianthus (Excalibur, Echo and Mariachi) for 12, 24, 36 and 48 days at temperatures of 5, 10 and 15°C, in the production and quality of buds, making this technology feasible to large-scale production. During cultivation it was observed that the lower the temperature and higher the vernalization period, the lower the cycle and the greater the number of plants induced to flowering for all three cultivars, and those are important features in the context of flower production in a commercial scale. The seeds subjected to vernalization originated plants that produce flower stems within the standards required by the market, showing that vernalization was efficient to induce flowering without affecting the quality of the buds. To produce lisianthus as a cut flower of quality, it is recommended seed vernalization of Mariachi and Echo cultivars for 24 days at 5°C and Excalibur for 36 days at 5°C.

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