Characterization of kinesin-like proteins in silkworm posterior silkgland cells

Kinesins 是涉及各种各样的细胞内部的运输的基于微导管的马达。神经原，鞭打的房间，和颜料房间传统地被用作模型系统学习 kinesins 的细胞的功能。这里，我们报导蚕以后的 silkgland (PSG ) ，为细胞内部的运输和丝心蛋白的有效分泌物与一个广泛的 endomembrane 系统专业化房间，，为 kinesin 学习的一个新奇模型。在 PSG 房间调查驾驶 kinesin 的细胞内部的运输，我们克隆五蚕象 kinesin 一样蛋白质(KLP ) ， BmKinesin-1， BmKinesin-6， BmKinesin-7， BmKinesin-13，和 BmKinesin-14A。我们由相对即时 PCR 并且西方的弄污决定了他们的表示模式。Immunofluorescence 显微镜学与微导管验证了他们的 colocalization。由联合下拉试金， LC-MS/MS，和西方的弄污分析，我们在 PSG，包括的包含丝心蛋白的小粒和联系 exuperantia 的 ribonucleoprotein (RNP ) 建筑群识别了 BmKinesin-1 的许多潜在的货物。而且， BmKinesin-13 overexpression 在 BmN 细胞破坏了微导管网络，它与在在另外的有机体调整微导管动力学的 Kinesin-13 的一个角色一致。根据这些结果，我们断定 PSG 可能在阐明有优点在能分泌的纸巾的细胞内部的运输的机制并且能为 kinesin 研究担任一个潜在的模型。

Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy

Technology development has always been one of the forces driving breakthroughs in biomedical research. Since the time of Thomas Morgan, Drosophilists have, step by step, developed powerful genetic tools for manipulating and functionally dissecting the Drosophila genome, but room for improving these technologies and developing new techniques is still large, especially today as biologists start to study systematically the functional genomics of different model organisms, including humans, in a high-throughput manner. Here, we report, for the first time in Drosophila, a rapid, easy, and highly specific method for modifying the Drosophila genome at a very high efficiency by means of an improved transcription activator-like effector nuclease （TALEN） strategy. We took advantage of the very recently developed ＂unit assembly＂ strategy to assemble two pairs of specific TALENs designed to modify the yellow gene （on the sex chromosome） and a novel autosomal gene. The mRNAs of TALENs were subsequently injected into Drosophila embryos. From 31.2% of the injected Fo fertile flies, we detected inheritable modification involving the yellow gene. The entire process from construction of specific TALENs to detection of inheritable modifications can be accomplished within one month. The potential applications of this TALEN-mediated genome modification method in Drosophila are discussed.

AtFH8 Is Involved in Root Development under Effect of Low-Dose Latrunculin B in Dividing Cells

Formins have been paid much attention for their potent nucleating activity. However, the connection between the in vivo functions of AtFHs （Arabidopsis thaliana formin homologs） and their effects on actin organization is poorly understood, in this study, we characterized the bundling activity of AtFH8 in vitro and in vivo. Biochemical analysis showed that AtFH8（FH1FH2） could form dimers and bundle preformed actin filaments or induce stellar structures during actin polymerization. Expression of truncated forms of AtFH8 and immunolocalization analysis showed that AtFH8 localized primarily to nuclear envelope in interphase and to the new cell wall after cytokinesis, depending primarily on its N-terminal transmembrane domain. GUS histochemical staining showed AtFH8 was predominantly expressed in Arabidopsis root meristem, vasculature, and outgrowth points of lateral roots. The primary root growth and lateral root initiation of atfh8 could be decreased by latrunculin B （LatB）. Analysis of the number of dividing cells in Arabidopsis root tips showed that much fewer dividing cells in Lat B-treated atfh8 plants than wild-type plants, which indicates that AtFH8 was involved in cell division. Actin cytoskeleton in root meristem of atfh8-1 was more sensitive to LatB treatment than that of wild-type. Altogether, our results indicate that AtFH8 is an actin filament nucleator and bundler that functions in cell division and root development.

STING-mediated DNA sensing in cancer immunotherapy

While STING(STimulator of INterferon Genes) has been shown to be essential for cytosolic DNA-triggered innate immune activation, accumulated evidence obtained from various studies suggested that an intrinsic relevance of STING-associated signaling in tumorigenesis can be observed. Also, several clinical trials using immunostimulatory adjuvants, particularly agonistic as well as non-agonistic ligands for STING, have revealed their therapeutic potential not only as vaccine adjuvants but also as anti-tumor agents. However, cases have also been reported where the involvement of STING shows a protective role in tumor growth. Here we summarize recent findings that have pointed towards the STING pathway as an innate immune sensing mechanism driving type I interferon production in the tumor context. Better understanding of this pathway can guide further development of novel immunotherapeutic strategies in the treatment of cancer.

Reverse Genetic Approaches in Zebrafish

Zebrafish（Danio rerio） is a well-established vertebrate animal model.A comprehensive collection of reverse genetics tools has been developed for studying gene function in this useful organism.Morpholino is the most widely used reagent to knock down target gene expression post-transcriptionally.For a long time,targeted genome modification has been heavily relied on large-scale traditional forward genetic screens,such as ENU（N-ethyl-N-nitrosourea） mutagenesis derived TILLING（Targeting Induced Local Lesions IN Genomes） strategy and pseudo-typed retrovirus mediated insertional mutagenesis.Recently,engineered endonucleases,including ZFNs（zinc finger nucleases） and TALENs（transcription activator-like effector nucleases）,provide new and efficient strategies to directly generate site-specific indel mutations by inducing double strand breaks in target genes.Here we summarize the major reverse genetic approaches for loss-of-function studies used and emerging in zebrafish,including strategies based on genome-wide mutagenesis and methods for site-specific gene targeting.Future directions and expectations will also be discussed.