COBRE: CELL CULTURE AND MOLECULAR BIOLOGY: CORE B

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 核心B：细胞和分子生物学核心 核心B应满足Cobre PI的所有细胞培养、分子生物学和荧光激活细胞分类(FACS)需求，这些PI在自己的实验室中不经常使用这些方法。 核心B可以提供来自胚胎皮质或脊髓的干细胞和/或血统受限的前体细胞。它们可以来自结构性表达内源标记的胎儿，如绿色荧光蛋白(GFP)或人胎盘碱性磷酸酶(HPLP)，或者它们可以被外源标记或诱导表达转基因，如下所述。 CoreB可以提供逆转录病毒、腺病毒和慢病毒载体，这些载体可以表达研究人员需要的任何cdna。此外，我们还有可调节的逆转录病毒(Teton和TetOFF)，它们在体外和体内都对多西环素有反应。 核心B已经开发出表达siRNA盒的慢病毒，我们可以针对任何Cobre研究人员想要沉默的mRNA。 核心B还开发了核糖核酸酶保护分析，可以根据所有研究人员的需要进行修改。 Core B可以提供定量实时聚合酶链式反应的专业技术，以评估任何感兴趣的基因的mRNA水平。 所有这些试剂在分散到Cobre PI之前都经过了核心的表征，以确保质量控制。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. CORE B: Cell and Molecular Biology CORE CORE B shall meet all of the cell culture, molecular biology, and fluorescence-activated cell sorting (FACS) needs of the COBRE PIs who are not routinely using these methods in their own laboratories. CORE B can provide stem and/or lineage restricted precursor cells derived from embryonic cortex or spinal cord. These can be from fetuses that constitutively express endogenous labels such as green fluorescent protein (GFP) or human placental alkaline phosphatase (hPLP) or they can be exogenous labeled or induced to express transgenes as described below. CORE B can provide retroviral, adenoviral, and lentiviral vectors that express whatever cDNA the Investigators' need. In addition, we have regulatable retroviruses (TetON and TetOFF) that are responsive to doxycyline, both in vitro and in vivo. CORE B has developed lentiviruses that express siRNA cassettes that we can target any mRNA that the COBRE Investigators' want silenced. CORE B has also developed RNAse protection assays that can be modified to all Investigators' needs. CORE B can provide expertise in quantiative real time PCR to assess mRNA levels for any gene of interest. All of these reagents have been characterized by the CORE prior to their dispersal to the COBRE PIs to ensure quality control.