生物谷会议频道

Fluidigm BioMark-陈巍学基因（22）

Fluidigm公司出品的BioMark系统，是一个基于微流控的，高通量的实时定量PCR系统。它可以高效、快速地对多个样本的、多个基因的表达量进行检测，也可以对多个样本的、多个SNP位点进行分型。同时它还可以大量地节约试剂、人工、实验时间。

本视频介绍了BioMark系统的工作原理，和其优势、特点。

2015-08-27 课时：8分钟

秦正红：DRAM1 regulates autophagy flux and Bid-me<font>di</font>ated cell death via lysosomes

秦正红：DRAM1 regulates autophagy flux and Bid-mediated cell death via lysosomes

秦正红，博士，教授，神经药理专业博士生导师。1994年在美国宾州医学院研究生院获博士学位，先后在美国国家卫生研究院(NIH)及麻省总医院和哈佛大学医学院从事研究工作。2003年从哈佛大学引进，现为苏州大学医学部基础医学与生物科学学院科研中心实验室主任，中国药理学会生化药理学专业委员会委员，中国药理学会神经药理学专业委员会委员，美国神经科学学会会员。

Damage-regulated autophagy modulator1 (DRAM1), a novel TP53 target gene, is an evolutionarily conserved lysosomal protein and plays an essential role in TP53-dependent autophagy activation and apoptosis (Crighton et al, 2006). However, the mechanisms by which DRAM1 promotes autophagy and apoptosis are not clear. 3-Nitropropionic acid (3-NP) is an inhibitor of mitochondrial respiratory complex II. Intrastriatal administration of 3-NP produces neuropathology resemble to Huntington disease. 3-NP-induced neuronal death was involved in autophagy and apoptosis. In vitro studies with 3-NP in TP53 wt and null cells, 3-NP or CCCP increased the protein levels of DRAM1 in a TP53-dependent or independent manner. DRAM1 induction contributed to 3-NP-induced autophagy activation. Knock-down of DRAM1 with siRNA inhibited the activity of V-ATPase, acidification of lysosomes and activation of lysosomal proteases. Knock-down of DRAM1 reduced the clearance of autophagososmes.

3-NP also induced a transcription independent upregulation of BAX protein levels. Knock-down of DRAM1 suppressed the increase in BAX levels. Co-immunoprecipitation and pull-down studies revealed an interaction of DRAM1 and BAX protein. Stably expression of exogenous DRAM1 increased the half-life of BAX. Upregulation of DRAM1 recruited BAX to lysosomes and induced cathepsin B-dependent cleavage of Bid and cytochrome c release. Knockdown of DRAM1, BAX or inhibition of lysosomal enzymes reduced 3-NP-induced cytochrome c release and cell death.

These data suggest that DRAM1 plays important roles in regulating autophagy flux and apoptosis. DRAM1 promotes autophagy flux through a mechanism involves activation of V-ATPase and enhances the acidification of lysosomes. DRAM1 promotes apoptosis via a mechanism involving recruitment of BAX to lysosomes to trigger cathepsin B-mediated Bid cleavage.

2015-09-30 课时：39分钟

GE：Dharmacon E<font>di</font>t-R基因编辑平台——即开即用，任性编辑！

GE：Dharmacon Edit-R基因编辑平台——即开即用，任性编辑！

CRISPR-Cas9基因编辑系统正以前所未有的速度席卷科研界。自从GE Dharmacon推出Edit-R CRISPR-Cas9系统以来，其"无需克隆，任性编辑"的特点广受好评。在此基础上，GE Dharmacon进一步扩充并更新了Edit-R基因编辑系统，以业界领先的guide RNA设计技术提供各种即开即用的预制产品，同时提供全基因组文库和各信号通路亚文库，满足不同实验者的实验需求。

2016-04-05 课时：49分钟

Stability of Morphogen Gra<font>di</font>ents & Movement of Molecules

Stability of Morphogen Gradients & Movement of Molecules

In my second lecture I describe experiments using EGFP tagged Bicoid to follow Bcd gradient establishment in living embryos, and to test various aspects of the simple model. Despite continuous synthesis of new Bcd protein at the anterior end of the egg, we find that the concentration of Bcd in nuclei at any given point along the anterior posterior axis is constant over time and is reproducible from embryo to the next. This reproducibility means that the gradient is sufficiently robust to provide positional information and thus can accurately direct gene activities. One the other hand, quantitative imaging experiments point to several features of the gradient that are hard to explain - how target genes activated by Bcd distinguish relatively subtle differences in low concentrations, and how Bcd molecules move from the anterior site of their synthesis to the site of their transcriptional activity. See more at http://www.ibioseminars.org

2016-04-21 课时：38分钟