COBRE: CELL CULTURE AND MOLECULAR BIOLOGY: CORE B

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 核心B：细胞和分子生物学核心 CORE 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选（FACS）需求，COBRE PI在其自己的实验室中不经常使用这些方法。 CORE B可以提供来源于胚胎皮层或脊髓的干细胞和/或谱系限制的前体细胞。 这些可以来自组成型表达内源性标记如绿色荧光蛋白（GFP）或人胎盘碱性磷酸酶（hPLP）的胎儿，或者它们可以是外源性标记的或诱导表达如下所述的转基因。 CORE B可以提供逆转录病毒、腺病毒和慢病毒载体，表达研究者需要的任何cDNA。 此外，我们有可调节的逆转录病毒（TetON和TetOFF），在体外和体内都对多西环素有反应。 CORE B已经开发出表达siRNA盒的慢病毒，我们可以靶向COBRE研究人员想要沉默的任何mRNA。 CORE B还开发了RNA酶保护试验，可根据所有研究者的需求进行修改。 CORE B可以提供定量真实的时间PCR的专业知识，以评估任何感兴趣基因的mRNA水平。 所有这些试剂在分散到COBRE PI之前都已由CORE进行了表征，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.