| Advances in Proximity Ligation in situ Hybridization (PLISH)
|
|
Author:
Date:
2020-11-05
[Abstract] Understanding tissues in the context of development, maintenance and disease requires determining the molecular profiles of individual cells within their native in vivo spatial context. We developed a Proximity Ligation in situ Hybridization technology (PLISH) that enables quantitative measurement of single cell gene expression in intact tissues, which we have now updated. By recording spatial information for every profiled cell, PLISH enables retrospective mapping of distinct cell classes and inference of their in vivo interactions. PLISH has high sensitivity, specificity and signal to noise ratio. It is also rapid, scalable, and does not require expertise in molecular biology so it can be easily adopted by basic and clinical researchers.
[摘要] [摘要]在发育，维持和疾病的背景下了解组织需要确定单个细胞在其天然体内空间范围内的分子谱。我们开发了一种邻近连接原位杂交技术（PLISH），该技术能够定量测量完整组织中单细胞基因的表达，现已更新。通过记录每个分析细胞的空间信息，PLISH可以回顾性绘制不同细胞类别并推断其体内 互动。PLISH具有很高的灵敏度，特异性和信噪比。它也快速，可扩展，并且不需要分子生物学方面的专门知识，因此基础和临床研究人员可以轻松地采用它。
[背景技术]我们最近开发了一种复用原位称为PLISH（邻位连接杂交技术原位杂交）（Nagendran等人，2018）。PLISH与其他现有的空间转录组学技术不同，因为它结合了高性能，快速多路复用，低成本和技术简单性（Wilbrey -Clark等人，2020年）。可以通过自动计算完整的冷冻或石蜡包埋组织中单细胞表达图谱来分析PLISH结果，它与同时进行的免疫染色兼容。
...
|
|
|
| Using RNA Sequencing and Spike-in RNAs to Measure Intracellular Abundance of lncRNAs and mRNAs
|
|
Author:
Date:
2020-10-05
[Abstract] Long noncoding RNAs (lncRNAs) play essential roles in normal physiology and in disease but their mechanisms of action can be challenging to identify. For mechanistic studies, it is often useful to know a lncRNA’s intracellular abundance, i.e., approximately how many molecules of the lncRNA are present in a typical cell of a cell-type of interest. At least two approaches have been used to approximate lncRNA intracellular abundance: single-molecule sensitivity RNA fluorescence in situ hybridization (smFISH) and single-gene, calibrated reverse-transcription followed by quantitative PCR (RT-qPCR). However, like all experimental approaches, these methods have their limitations. smFISH, when analyzed using diffraction-limited microscopy, can underestimate intracellular ...
[摘要] [摘要]长非编码RNA（lncRNA）在正常生理和疾病中起着至关重要的作用，但其作用机理可能难以鉴定。对于机理研究，了解lncRNA的胞内丰度（即在目标细胞类型的典型细胞中大约存在多少个lncRNA分子）通常很有用。至少两种方法已用于估算lncRNA细胞内丰度：单分子敏感性RNA荧光原位杂交（smFISH ...
|
|
|
| Preparation and Characterization of Ginger Lipid-derived Nanoparticles for Colon-targeted siRNA Delivery
|
|
Author:
Date:
2020-07-20
[Abstract] Synthetic nanoparticle-based drug delivery system is widely known for its ability to increase the efficacy and specificity of loaded drugs, but it often suffers from relatively higher immunotoxicity and higher costs as compared to traditional drug formulations. Contrarily, plant-derived nanoparticles appear to be free from these limitations of synthetic nanoparticles; they are naturally occurring biocompatible vesicles that do not generate immunotoxicity and are easy to obtain. Additionally, lipids isolated from plant-derived nanoparticles have shown the capability of assembling themselves to spherical nano-sized liposomal particles. Herein, we employ lipids extracted from ginger-derived nanoparticles and load them with therapeutic siRNA (CD98-siRNA) to create CD98-siRNA/ginger-lipid ...
[摘要] [摘要 ]基于合成纳米颗粒的药物递送系统以其增加所载药物的功效和特异性的能力而广为人知，但是与传统药物制剂相比，它经常遭受相对较高的免疫毒性和较高的成本。相反，植物来源的纳米粒子似乎不受合成纳米粒子的这些限制；它们是天然存在的生物相容性囊泡，不会产生免疫毒性并且易于获得。另外，从植物来源的纳米颗粒分离的脂质已显示出将其自身组装成球形纳米尺寸脂质体颗粒的能力。在本文中，我们采用从生姜纳米颗粒中提取的脂质，并用治疗性siRNA（CD98-siRNA）加载脂质，以创建CD98-siRNA /生姜脂质纳米颗粒。CD98-siRNA /姜脂脂质纳米颗粒的表征显示它们呈球形，直径约为189.5 nm。纳米颗粒的表面ζ电势在-18.1至-18.4mV之间变化。此外，在最近的研究中，CD98-siRNA /姜脂纳米颗粒在右旋糖酐硫酸钠（DSS）诱发的结肠炎小鼠模型中显示出特定的结肠靶向能力和出色的抗炎功效。
[背景 ] 小干扰RNA（siRN As）是一种有前途的治疗剂，可以通过沉默异常上调的信使RNA（mRNA）来治疗各种疾病（Nikam and Gore ，2018）。尽管siRNA的有效性，安全有效地将siRNA递送至治疗靶标仍然是一项艰巨的任务（Tatiparti et al ...
|
|
|