Molecular Biology


    Protocols in Current Issue
    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: 185, 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: 549, 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: 1863, 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: 3184, 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: 5109, 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: 4341, 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: 5268, 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: 6173, Q&A: 0
    [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 ...
    Genome-Wide siRNA Screen for Anti-Cancer Drug Resistance in Adherent Cell Lines
    Authors:  Elza C. de Bruin, Ming Jiang, Michael Howell and Julian Downward, date: 05/20/2015, view: 7166, Q&A: 0
    [Abstract] The expression of genes is frequently manipulated in cell lines to study their cellular functions. The use of exogenous small Interfering RNAs (siRNAs) is a very efficient technique to temporarily downregulate the expression of genes of interest [reviewed by Hannon and Rossi (2004)]. A genome-wide siRNA library allows the user to study both the ...
    In vitro Reconstitution Assay of miRNA Biogenesis by Arabidopsis DCL1
    Authors:  Tian Wang, Claudia Castillo-González, Lan You, Rui Li, Liwei Wen, Hongliang Zhu and Xiuren Zhang, date: 04/20/2015, view: 4848, Q&A: 0
    [Abstract] microRNAs (miRNAs) are small non-coding RNAs, regulating most if not all, biological processes in eukaryotic organisms. miRNAs are initially processed from primary transcripts (pri-miRNAs) to produce miRNA precursors (pre-miRNAs), that are further processed into miRNA and its complementary strands (miRNA/*). In Arabidopsis, and possibly other ...

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