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分子生物学

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    现刊
    In vitro Assays for Eukaryotic Leading/Lagging Strand DNA Replication
    真核先导链/后随链DNA复制的体外分析
    作者:Grant Schauer, Jeff Finkelstein and Mike O’Donnell日期:09/20/2017,浏览量:4,Q&A: 0
    The eukaryotic replisome is a multiprotein complex that duplicates DNA. The replisome is sculpted to couple continuous leading strand synthesis with discontinuous lagging strand synthesis, primarily carried out by DNA polymerases ε and δ, ...
    Method for Multiplexing CRISPR/Cas9 in Saccharomyces cerevisiae Using Artificial Target DNA Sequences
    酿酒酵母中使用人工靶DNA序列进行多重CRISPR/ Cas9的方法
    作者:Rachael M. Giersch and Gregory C. Finnigan日期:09/20/2017,浏览量:4,Q&A: 0
    Genome manipulation has become more accessible given the advent of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) editing technology. The Cas9 endonuclease binds a single stranded (single guide) RNA (sgRNA) fragment that ...
    Construction of a Single Transcriptional Unit for Expression of Cas9 and Single-guide RNAs for Genome Editing in Plants
    构建共表达Cas9和sgRNAs的单一独立转录单元用于植物基因组编辑
    作者:Xu Tang, Zhaohui Zhong, Xuelian Zheng and Yong Zhang日期:09/05/2017,浏览量:145,Q&A: 0
    The CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein9 (Cas9) is a simple and efficient tool for genome editing in many organisms including plant and crop species. The sgRNAs of the CRISPR/Cas9 system are ...
    Protocol for Establishing a Multiplex Image-based Autophagy RNAi Screen in Cell Cultures
    在细胞培养中建立基于多重图像的自噬RNAi筛选方法
    作者:Jennifer Jung and Christian Behrends日期:09/05/2017,浏览量:118,Q&A: 0
    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 ...
    Using CRISPR-ERA Webserver for sgRNA Design
    使用网络服务器CRISPR-ERA设计sgRNA
    作者:Honglei Liu, Xiaowo Wang and Lei S. Qi日期:09/05/2017,浏览量:99,Q&A: 0
    The CRISPR-Cas9 system is emerging as a powerful technology for gene editing (modifying the genome sequence) and gene regulation (without modifying the genome sequence). Designing sgRNAs for specific genes or regions of interest is indispensable to ...
    In vitro Assays for Eukaryotic Leading/Lagging Strand DNA Replication
    真核先导链/后随链DNA复制的体外分析
    作者:Grant Schauer, Jeff Finkelstein and Mike O’Donnell日期:09/20/2017,浏览量:4,Q&A: 0
    [Abstract] The eukaryotic replisome is a multiprotein complex that duplicates DNA. The replisome is sculpted to couple continuous leading strand synthesis with discontinuous lagging strand synthesis, primarily carried out by DNA polymerases ε and δ, respectively, along with helicases, polymerase α-primase, DNA sliding clamps, clamp loaders and many other ...
    Method for Multiplexing CRISPR/Cas9 in Saccharomyces cerevisiae Using Artificial Target DNA Sequences
    酿酒酵母中使用人工靶DNA序列进行多重CRISPR/ Cas9的方法
    作者:Rachael M. Giersch and Gregory C. Finnigan日期:09/20/2017,浏览量:4,Q&A: 0
    [Abstract] Genome manipulation has become more accessible given the advent of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) editing technology. The Cas9 endonuclease binds a single stranded (single guide) RNA (sgRNA) fragment that recruits the complex to a corresponding genomic target sequence where it induces a double stranded ...
    Construction of a Single Transcriptional Unit for Expression of Cas9 and Single-guide RNAs for Genome Editing in Plants
    构建共表达Cas9和sgRNAs的单一独立转录单元用于植物基因组编辑
    作者:Xu Tang, Zhaohui Zhong, Xuelian Zheng and Yong Zhang日期:09/05/2017,浏览量:145,Q&A: 0
    [Abstract] The CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein9 (Cas9) is a simple and efficient tool for genome editing in many organisms including plant and crop species. The sgRNAs of the CRISPR/Cas9 system are typically expressed from RNA polymerase III promoters, such as U6 and U3. In many transformation events, ...
    Protocol for Establishing a Multiplex Image-based Autophagy RNAi Screen in Cell Cultures
    在细胞培养中建立基于多重图像的自噬RNAi筛选方法
    作者:Jennifer Jung and Christian Behrends日期:09/05/2017,浏览量:118,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 ...
    Using CRISPR-ERA Webserver for sgRNA Design
    使用网络服务器CRISPR-ERA设计sgRNA
    作者:Honglei Liu, Xiaowo Wang and Lei S. Qi日期:09/05/2017,浏览量:99,Q&A: 0
    [Abstract] The CRISPR-Cas9 system is emerging as a powerful technology for gene editing (modifying the genome sequence) and gene regulation (without modifying the genome sequence). Designing sgRNAs for specific genes or regions of interest is indispensable to CRISPR-based applications. CRISPR-ERA (http://crispr-era.stanford.edu/ ...
    Assessment of Modulation of Protein Stability Using Pulse-chase Method
    使用脉冲追踪法评估蛋白质稳定性的调节
    作者:Mohamed Elgendy日期:08/20/2017,浏览量:85,Q&A: 0
    [Abstract] Pulse-chase technique is a method widely used to assess protein or mRNA stability. The principle of pulse-chase relies on labeling proteins or mRNA produced during a short period of time called ‘pulse’ and then following the rate of disappearance of those labeled proteins over a period of time called ‘chase’. This technique thus allows ...
    A High-throughput Assay for mRNA Silencing in Primary Cortical Neurons in vitro with Oligonucleotide Therapeutics
    体外使用寡核苷酸治疗药物对原代皮质神经元mRNA沉默的高通量测定
    [Abstract] Primary neurons represent an ideal cellular system for the identification of therapeutic oligonucleotides for the treatment of neurodegenerative diseases. However, due to the sensitive nature of primary cells, the transfection of small interfering RNAs (siRNA) using classical methods is laborious and often shows low efficiency. Recent progress in ...
    TUNEL Assay to Assess Extent of DNA Fragmentation and Programmed Cell Death in Root Cells under Various Stress Conditions
    各种胁迫条件下根细胞DNA片段化和程序性细胞死亡的TUNEL法评估
    作者:Amit K. Tripathi, Ashwani Pareek and Sneh Lata Singla-Pareek日期:08/20/2017,浏览量:84,Q&A: 0
    [Abstract] DNA damage is one of the common consequences of exposure to various stress conditions. Different methods have been developed to accurately assess DNA damage and fragmentation in cells and tissues exposed to different stress agents. However, owing to the presence of firm cellulosic cell wall and phenolics, plant cells and tissues are not easily ...
    Establishment of a Human Cell Line Persistently Infected with Sendai Virus
    持续感染仙台病毒的人细胞系的建立
    作者:Christopher Coakley, Cara Peter, Stephanie Fabry and Saurabh Chattopadhyay日期:08/20/2017,浏览量:75,Q&A: 0
    [Abstract] Interferon regulatory transcription factor 3 (IRF3) is a transcription factor that upon activation by virus infection promotes the synthesis of antiviral genes, such as the interferons (Hiscott, 2007). In addition to inducing genes, IRF3 triggers antiviral apoptosis by RIG-I-like receptor-induced IRF3 mediated pathway of apoptosis (RIPA), which is ...
    Antisense Oligonucleotide-mediated Knockdown in Mammary Tumor Organoids
    乳腺肿瘤类器官中反义寡核苷酸介导的基因敲低技术
    作者:Sarah D. Diermeier and David L. Spector日期:08/20/2017,浏览量:77,Q&A: 0
    [Abstract] Primary mammary tumor organoids grown in 3D are an excellent system to study tumor biology. They resemble the organization and physiology of native epithelia more closely than cancer cell lines grown in 2D, and additionally model interactions with the ECM (Boj et al., 2015; Clevers, 2016; Shamir and Ewald, 2014). Mammary tumor organoids ...