Dendrimers and Gene Delivery
Zahoor Ahmad Parry, Rajesh Pandey in Dendrimers in Medical Science, 2017
Gene therapy is a proven tool to tackle many suitable selected stubborn diseases such as cancer and genetic/hereditary disorders in which conventional treatments may be ineffective. Overall, dendrimer-mediated delivery of therapeutic nucleic acids including silencing RNA (siRNA) definitely is to be considered a promising approach. The unique features of dendrimeric systems including the ease of surface functionalization, enables the crafting of truly versatile nanodevices for drug delivery applications. However, careful addressing of the recognized current limitations that include non-specific cytotoxicity observed with higher generation-dendrimers, release kinetics and fast clearance would open up new vistas for dendrimers as state of the art gene delivery carriers. A family of triazine dendrimers with varying core flexibility, generation as well as surface functionality was assessed for its potential to condense and efficiently deliver a luciferase targeting siRNA aimed at reporter gene knockdown. The carbosilane dendrimers, another new class, has been employed for siRNA delivery.
The Preparation and Properties of Immobilized Firefly Luciferase for Use in the Detection of Microorganisms
Wilfred H. Nelson in Physical Methods for Microorganisms Detection, 1991
This chapter analyzes the results of the immobilizations, to introduce some unpublished data on the methods available for immobilization of luciferase, and to discuss the potential of these methods. Immobilized iuciferase preparations generally have a longer transient period before maximum light emission is achieved. The immobilized luciferase produced from the doubly activated membrane showed a small increase in activity but this was thought to be too low relative to the increased effort needed to obtain it. A much more sensitive process involves the luminescence reaction catalyzed by firefly luciferase. This can be used for the analysis of Adenosine triphosphate (ATP) down to femtomolar concentrations and with a linear response up to micromolar concentrations. Many methods suggested for the analysis of ATP. Such methods have received close attention from microbiologists as the concentration of viable microorganisms is closely related to that of the ATP they contain and the endogenous ATP may be released by simply lysing the cells using various reagents.
Bioluminescence
Margarida M. Barroso, Xavier Intes in Imaging from Cells to Animals, 2020
This chapter focuses on bioluminescent imaging in vivo and how the development of modern luciferase systems enable this goal. A diverse set of organisms naturally emit light using endogenous chemical pathways. Modern biotechnology has exploited this luminescent biochemistry to extend our understanding of the natural world, dissect disease processes, and generate novel medications. In contrast to the Fluc system discussed, the bacterial lux system enables an alternative imaging approach, whereby the pathway producing the luciferin is genetically encoded and delivered into the target cells alongside the luciferase itself. The luciferase reactions discussed have been controlled genetically, in which the presence of the luciferase is a result of promoter activation or defined by the start of luciferin administration.
Bioluminescence assays: multicolor luciferase assay, secreted luciferase assay and imaging luciferase assay
Published in Expert Opinion on Drug Discovery, 2010
Yoshihiro Nakajima, Yoshihiro Ohmiya
Importance of the field: By selecting the most appropriate bioluminescence assay, the researcher can study the underlying molecular mechanisms of a physiological system and the effects of a drug throughout the body. Areas covered in this review: This review covers three luciferase assay systems: the multicolor luciferase assay, secreted luciferase assay and imaging luciferase assay. These assays are applied to drug screening in vitro, in cellulo and in vivo. What the reader will gain: Different solutions for reporter assay in vitro, in cellulo and in vivo are presented. A suitable bioluminescence system depending on the assay purpose is also discussed. Take home message: Bioluminescence is a manifold system based on the different types of luciferin and its luciferase. Namely, luciferin catalyzed by corresponding luciferases resulted in the production of different color lights. We must understand the manifold mechanisms of bioluminescence reaction.
Upregulation of miR-92a contributes to blocking goblet cell metaplasia by targeting MUC5AC in asthma
Published in Journal of Receptors and Signal Transduction, 2020
Jihuan Dai, Bo Ma, Xiaolin Wen, Zhouxin Yang, Yingxing Yue
As a chronic airway disease, asthma has two characteristics, tissue remodeling and airway inflammation. This research focused on miR-92a to explore how it works in asthma. We revealed that the expressions of miR-92a were decreased in both serum and lung tissues from ovalbumin-induced asthma mouse. Bioinformatics analysis, quantitative polymerase chain reaction (qPCR) and dual luciferase assay revealed that miR-92a targets MUC5AC, which was linked to mucus hypersecretion in the pulmonary tracts. By injecting miR-92a-mimics into the trachea, both the airway hyper-reactivity and airway inflammation can be alleviated in an asthma mouse model which is induced by ovalbumin. Moreover, the goblet cell phenotype of asthmatic mice is significantly reduced by the action of miR-92a. Furthermore, miR-92a blocked interleukin (IL)-13-induced MUC5AC luciferase activity in 16HBE. Together, upregulation of miR-92a expression in asthmatic mice plays a role in blocking goblet cell metaplasia by targeting MUC5AC, and thus in the treatment of chronic airway diseases, miR-92a can prevent epithelial remodeling, which is a reasonable method.
Dynamic changes in miR-124 levels in patients with acute cerebral infarction
Published in International Journal of Neuroscience, 2019
Mei Sun, Xiaoxia Hou, Guang Ren, Yangyang Zhang, Hong Cheng
Objective: To investigate the changes in serum miR-124 levels in patients with acute cerebral infarction (ACI) and elucidate the underlying mechanism by a dynamic monitor. Methods: Fifty-four patients with ACI and 51 healthy controls were included in our study. Baseline characteristics and blood samples were collected for further analysis. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the serum miR-124 levels. The dual-luciferase reporter assay was used to evaluate the effect of miR-124 on iASPP, a protein that inhibits apoptosis stimulating proteins in the p53 family. Results: Compared with normal controls, the miR-124 levels in the ACI group rapidly decreased at phase 1 (within 24 h after ischemia) (p
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