SUBCONTRACT TO WISCONSIN STEM CELL RESEARCH CENTER

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。通过同源重组将新霉素抗性或绿色荧光蛋白等可选择标记引入组织特异性基因。我们通过同源重组将绿色荧光蛋白和嘌呤霉素导入到位于染色体13q34上的hSox1中。共获得8个克隆。当这些hSox1敲入细胞进行神经分化时，RT-PCR检测到低水平的EGFP mRNA表达。我们计划优化神经分化方案并纯化hSox1细胞，以确定其发育潜力并分析其基因表达。这项研究使用了WNPRC干细胞资源和联邦批准的人类ES细胞系。

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. To introduce a selectable marker such as neomycin resistance or EGFP by homologous recombination into a tissue specific gene. We introduced EGFP and puromycin into hSox1 located on chromosome 13q34 via homologous recombination. Eight clones were obtained. When these hSox1 knock in cells underwent neural differentiation low levels of EGFP mRNA expression was observed via RT-PCR. We plan to optimize the neural differentiation protocol and purify hSox1 cells to determine their developmental potential and analyze their gene expression. This research uses WNPRC Stem Cell Resources and federally approved human ES cell lines.