RNA interference —RNA —Molecular Biology

RNA Interference Method for Gene Function Analysis in the Japanese Rhinoceros Beetle Trypoxylus dichotomus

Authors: Kazuki Sakura, Shinichi Morita and Teruyuki Niimi, date: 04/20/2022, view: 721, Q&A: 0

[Abstract]

In the Japanese rhinoceros beetle Trypoxylus dichotomus, various candidate genes required for a specific phenotype of interest are listed by next-generation sequencing analysis. Their functions were investigated using RNA interference (RNAi) method, the only gene function analysis tool for T. dichotomus developed so far. The summarized procedure

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In vivo CD40 Silencing by siRNA Infusion in Rodents and Evaluation by Kidney Immunostaining

Authors: Miguel Hueso, Adrián Mallén, Elia Ripoll, Laura de Ramón, Nuria Bolaños, Cristian Varela, Jordi Guiteras, Javier Checa, Estanislao Navarro, Josep Maria Grinyo, Josep Maria Cruzado, Josep Maria Aran and Joan Torras, date: 05/20/2021, view: 6021, Q&A: 0

[Abstract]

The co-stimulatory molecule CD40 and its ligand CD40L play a key role in the regulation of immunological processes and are involved in the pathophysiology of autoimmune and inflammatory diseases. Inhibition of the CD40-CD40L axis is a promising therapy, and a number of strategies and techniques have been designed to hinder its functionality. Our

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pNP Transgenic RNAi System Manual in Drosophila

Authors: Fang Wang, Huan-Huan Qiao, Rong-Gang Xu, Jin Sun, Ruibao Zhu, Decai Mao and Jian-Quan Ni , date: 02/05/2019, view: 5332, Q&A: 0

[Abstract] Much of our knowledge about the mechanisms underlying biological processes relies on genetic approaches, whereby gene activity is reduced and the phenotypic consequences of perturbation are analyzed in detail. For functional genomic studies, a specific, systematic, and cost-effective manner is critical. Transgenic RNAi system is the top priority ...

Variable Dose Analysis: A Novel High-throughput RNAi Screening Method for Drosophila Cells

Authors: Katarzyna Sierzputowska, Chris R. Baxter and Benjamin E. Housden, date: 12/20/2018, view: 3719, Q&A: 0

[Abstract] Genetic screens are a powerful approach to identify previously uncharacterized genes involved in specific biological processes. Several technologies have been developed for high-throughput screens using reagents such as RNAi or CRISPR, and each approach is associated with specific advantages and disadvantages. Variable Dose Analysis (VDA), is an ...

Dual Fluorescence Reporter Based Analytical Flow Cytometry for miRNA Induced Regulation in Mammalian Cells

Authors: Nicolas Lemus-Diaz, Liezel Tamon and Jens Gruber, date: 09/05/2018, view: 9643, Q&A: 0

[Abstract] MicroRNA-induced gene regulation is a growing field in basic and translational research. Examining this regulation directly in cells is necessary to validate high-throughput data originated from RNA sequencing technologies. For this several studies employ luciferase-based reporters that usually measure the whole cell population, which comes with ...

Protocol for Establishing a Multiplex Image-based Autophagy RNAi Screen in Cell Cultures

Authors: Jennifer Jung and Christian Behrends, date: 09/05/2017, view: 7255, Q&A: 0

[Abstract] Autophagy is a recycling pathway, in which intracellular cargoes including protein aggregates and bacteria are engulfed by autophagosomes and subsequently degraded after fusion with lysosomes. Dysregulation of this process is involved in several human diseases such as cancer or neurodegeneration. Hence, advancing our understanding of how autophagy ...

Mungbean Yellow Mosaic India Virus (MYMIV)-infection, Small RNA Library Construction and Deep Sequencing for MicroRNA Identification in Vigna mungo

Authors: Anirban Kundu, Sujay Paul, Amita Pal and Genotypic Technology, date: 10/20/2016, view: 9629, Q&A: 0

[Abstract] This protocol describes small RNA library preparation from Vigna mungo total RNA followed by deep sequencing and analysis for microRNA identification.

Conjugation of Duplexed siRNN Oligonucleotides with DD-HyNic Peptides for Cellular Delivery of RNAi Triggers

Authors: Alexander S. Hamil, Khirud Gogoi and Steven F. Dowdy, date: 04/05/2016, view: 7606, Q&A: 0

[Abstract] Despite the great promise that short interfering RNA (siRNA) induced RNAi responses hold as a therapeutic modality, due to their size (~15 kDa) and high negative charge (Bumcrot et al., 2006), siRNAs have no bioavailability and require a delivery agent to enter cells (Figure 1). TAT peptide transduction domain (PTD) has been developed as ...

Virus-based MicroRNA Silencing

Authors: Jinping Zhao and Yule Liu, date: 01/20/2016, view: 9436, Q&A: 0

[Abstract] Virus-based microRNA silencing (VbMS) is a viable and prompt method to screen and characterize the function of microRNAs (miRNAs) in plants. The Tobacco rattle virus (TRV)-based VbMS method was originally developed by the Yule Liu's group (Sha et al., 2014) using miRNA target mimic (TM) methodology. Here, we describe the ...

Virus-induced Gene Silencing (VIGS) in Barley Seedling Leaves

Authors: Lokanadha R. Gunupuru, Shahin S. Ali, Fiona M. Doohan and Steven R. Scofield, date: 06/20/2015, view: 11580, Q&A: 1

[Abstract] Virus induced gene silencing (VIGS) is one of the most potent reverse genetics technologies for gene functional characterisation. This method exploits a dsRNA-mediated antiviral defence mechanism in plants. Using this method allows researchers to generate rapid phenotypic data in a relatively rapid time frame as compared to the generation of ...

Molecular Biology

Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat

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