SUBCONTRACT TO WISCONSIN STEM CELL RESEARCH CENTER

威斯康星干细胞研究中心的分包合同

• 批准号: 7716430
• 负责人: James Alexander Thomson
• 金额: $ 8.19万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-23 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7716430
• 关键词: Antibodies; Brachyury protein; Cell Differentiation process; Cell Separation; Computer Retrieval of Information on Scientific Projects Database; Condition; DNA; Funding; Genes; Grant; Institution; Magnetism; Mesoderm; Mesoderm Cell; Neomycin; Research; Research Personnel; Resistance; Resources; Source; Stem Cell Research; Stem cells; Techniques; Tissues; United States National Institutes of Health; Wisconsin; homologous recombination; human embryonic stem cell; human embryonic stem cell line; vector

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 identified culture conditions that allow human ES cells to efficiently differentiate into mesodermal lineages in defined conditions. Additionally we created a DNA vector that would tag, the brachyury gene with CD4, neomycin, and a soft tag sequence using homologous recombination. The successful tagging of the brachyury gene would allow us to purify early mesodermal cells with simple magnetic cell sorting technique. It would also enable us to study the functions of T gene in definite mesoderm commitment during human ES cell differentiation using a highly specific Soft tag antibody. The successful applications of Soft tag would also provide a universal tagging strategy to study the functions of many other genes, especially those lacking good antibodies. This research uses WNPRC Stem Cell Resources, the IS Division, and federally approved human ES cell lines.

这个子项目是许多研究子项目中的一个 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。 通过同源重组将选择标记如新霉素抗性或EGFP导入组织特异性基因。 我们确定了培养条件，使人ES细胞有效地分化成中胚层谱系在规定的条件。此外，我们创建了一个DNA载体，该载体将使用同源重组用CD 4、新霉素和软标签序列标记短尾畸形基因。Brachyury基因的成功标记将使我们能够用简单的磁性细胞分选技术纯化早期中胚层细胞。这也将使我们能够研究T基因的功能，在明确的中胚层定型在人ES细胞分化过程中，使用一个高度特异性的软标签抗体。Soft tag的成功应用也将为研究其他基因的功能，特别是那些缺乏良好抗体的基因的功能提供一种通用的标签策略。这项研究使用WNPRC干细胞资源，IS部门和联邦批准的人类ES细胞系。