China
Africa
Explore chapters and articles related to this topic
RNA-seq Analysis
Published in Altuna Akalin, Computational Genomics with R, 2020
Quantification of how much gene expression levels deviate from a baseline gives clues about which genes are actually important for, for instance, disease outcome or cell/tissue identity. The methods of detecting and quantifying gene expression have evolved from low-throughput methods such as the usage of a reporter gene with a fluorescent protein product to find out if a single gene is expressed at all, to high-throughput methods such as massively parallel RNA-sequencing that can profile -at single-nucleotide resolution- the abundance of tens of thousands of distinct transcripts encoded in the largest eukaryotic genomes.
Delivery of Genes Through the Lung Circulation
Published in Kenneth L. Brigham, Gene Therapy for Diseases of the Lung, 2020
David M. Rodman, Elizabeth G. Nabel
Gene delivery to the pulmonary circulation is feasible. Using present technology, isolation of the lung circulation with either a catheter-based or a surgical approach yields the highest expression. Both lipid and adenoviral vectors yield lung expression, though adenovirus produces higher efficiency. While systemic delivery utilizing lipid:DNA complexes appears to result primarily in endothelial cell expression, the higher levels of expression achieved with directed approaches result in expression in a variety of other lung cells, including alveolar epithelial cells. Of note, little expression in vascular smooth muscle cells has been detected in any study to date. The present studies still represent early stages of development of this technology. While a variety of techniques result in detectable reporter gene activity, it remains to be proven if these techniques provide sufficient efficiency to produce biological effects from genes of interest. Challenges for the future include development of more efficient vectors and improved cell targeting. Despite these limitations, pulmonary vascular gene transfer holds promise as a useful investigative and therapeutic tool.
Optimizing Reporter Gene Expression for Molecular Magnetic Resonance Imaging
Published in Shoogo Ueno, Bioimaging, 2020
Qin Sun, Frank S. Prato, Donna E. Goldhawk
Simply stated, a reporter gene encodes protein that can be reliably detected by some method of choice. If the reporter gene is constitutively expressed, it generates a constant label that can theoretically be monitored through all phases of the cell, including its proliferation and differentiation. If the reporter gene is selectively expressed, it imparts a signal that will only be detected when specific TF activity is present, including those that trigger a disease process like inflammation, fibrosis, or metastasis. Traditionally, “reporter gene expression” is a term coined to describe this type of selective gene expression. It is a powerful tool for understanding cellular phenotype and identifying what factors contribute to function and malfunction.
Strategies for targeting RNA with small molecule drugs
Published in Expert Opinion on Drug Discovery, 2023
Christopher L. Haga, Donald G. Phinney
As the name implies, cell-based RNA-small molecule screening systems are conducted within various cell lines. These systems have been used to identify small molecules capable of inhibiting the activity of noncoding RNAs such as miRNAs, translation by targeting 5’ untranslated regions (UTR) of mRNAs, and regulation of RNA splicing (Figure 3(c)). Cell-based systems employ the use of a reporter gene, typically a fluorescent gene such as GFP or a luminescent gene such as firefly luciferase, either upstream, downstream, or interrupting a target RNA gene. This reporter gene acts as a read-out for disruption of RNA function. Reporter constructs are transfected into a given cell line, incubated with compounds of interest, and analyzed by plate reader or cytometry for alterations in fluorescence or luminescence.
Monoclonal antibody as a targeting mediator for nanoparticle targeted delivery system for lung cancer
Published in Drug Delivery, 2022
Nasrul Wathoni, Lisa Efriani Puluhulawa, I Made Joni, Muchtaridi Muchtaridi, Ahmed Fouad Abdelwahab Mohammed, Khaled M. Elamin, Tiana Milanda, Dolih Gozali
The synthesis of functionally assembled supramolecular nanoparticles was performed for RNAi agent loading and tumor target treatment. The adamantane-grafted poly(ethylene glycol) molecule was modified with the specific binding ligand EGFR GE11 or the pH-sensitive fusogenic peptide GALA, which was then used for self-assembly with the cyclodextrin-grafted branched polyethyleneimine (CD-PEI), adamantane-grafted polyamidoamine dendrimer (Ad-PAMAM), and DNA. These nanoparticles showed that beneficial peptides can enhance targeted cell binding, internalization, and endosome release. Furthermore, it leads to enhanced reporter gene expression and effective target gene silencing. Systemic administration can effectively lower intratumoral VEGF protein levels, limit angiogenesis, and considerably suppress tumor development in A549 xenografts (Lu et al., 2020).
Approaching complexity: systems biology and ms-based techniques to address immune signaling
Published in Expert Review of Proteomics, 2020
Joseph Gillen, Caleb Bridgwater, Aleksandra Nita-Lazar
Reporter assays constitute the in vitro stimulation of a promoter and reporter gene pair to measure a response [2]. These assays can be extremely useful and are a robust method for high throughput screens of different drugs or compounds by the direct measurement of expressed gene products or the indirect changes in signal molecules regulated by genes. This is accomplished by transfecting a desired cell line to express a transcription factor activated promoter and a common reporter gene. The cells can then be stimulated to drive promoter activity. Ligand or agonist induction of the transcription factors leading to expression of the reporter gene. In the case of a luciferase reporter gene, luciferin, a luciferase substrate, can be added and the resulting luminescence measured as a corollary of stimulation. The unattractive variables of reporter assays tend to revolve around the limits in experimental design: reporter assays may take vast resources and long time periods to design and to target assays to transcription factors. Common reporter assays exploit the activity of reporter genes producing β-galactosidase, green fluorescent protein (GFP), and luciferase [2].