CAREER: Analyzing and engineering multi-gene cooperation to generate cell lines for therapeutic protein production

职业：分析和设计多基因合作以生成用于治疗性蛋白质生产的细胞系

• 批准号: 0846392
• 负责人: Clifford Wang
• 金额: $ 40万
• 依托单位: Stanford University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2014-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0846392&HistoricalAwards=false
• 关键词: CAREER; Analyzing; engineering; multi; gene

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).0846392WangIntellectual Merit: As cell biologists, geneticists, or engineers, one routinely over-expresses a gene in a cell, observes the cell, and compares its behavior to the parent. Yet multiple genes, not one, often determine cell phenotypes. One example is the sustained proliferation characteristic of immortalized mammalian cells. A mutation that changes the expression of a single gene or activity of a single protein rarely suffices to immortalize a cell; instead, multiple genes generally need to be mutated. This project investigates how multiple genes contribute to the phenotypes of immortalized cells: proliferation, apoptosis, senescence, and serum-independent growth. This project also aims to determine how different gene expression levels affect these phenotypes. In other words, the experiments address both combinatorial and dose effects. Broader Impact: This technology provides a powerful tool to generate cell lines with properties optimized for the production of therapeutic proteins. Using this experimental and analytical platform, this project will generate cell lines that exhibit high proliferation rates in low serum or serum-free media. Expression of various gene combinations will be evaluated for the ability to improve or create cell lines. Gene candidates include known oncogenes, tumor suppressor genes, and cell cycle regulators, as well as genes identified previously by retroviral mutagenesis in mice. The long-term goal is to generate human plasma B cell lines for the production of monoclonal antibodies.Faculty, graduate students, and undergraduates from the Department of Chemical Engineering at Stanford University will adopt a class of freshmen at Cupertino High School. This class is enrolled in a program named AVID (Advancement via Individual Determination). The program identifies students with poor class performance, but good attendance and minimal disciplinary problems. The program has no selection criteria based on race or gender, although the majority of students are economically disadvantaged, underrepresented minorities who will be the first in their families to attend college. The four-year program aims to help these students graduate from high school and go to college.

该奖项是根据2009年美国复苏和再投资法案（公法111-5）资助的。0846392 WangIntellectual Merit：作为细胞生物学家，遗传学家，或工程师，一个常规的过度表达细胞中的基因，观察细胞，并将其行为与父母进行比较。然而，多个基因，而不是一个，往往决定细胞表型。一个例子是永生化哺乳动物细胞的持续增殖特性。改变单个基因的表达或单个蛋白质的活性的突变很少足以使细胞永生;相反，多个基因通常需要突变。该项目研究多个基因如何促进永生化细胞的表型：增殖，凋亡，衰老和血清非依赖性生长。该项目还旨在确定不同的基因表达水平如何影响这些表型。换句话说，实验解决了组合效应和剂量效应。更广泛的影响：这项技术提供了一个强大的工具，用于生成具有优化特性的细胞系，用于生产治疗性蛋白质。利用这个实验和分析平台，该项目将产生在低血清或无血清培养基中表现出高增殖率的细胞系。将评价各种基因组合的表达改善或创建细胞系的能力。候选基因包括已知的癌基因、肿瘤抑制基因和细胞周期调节基因，以及先前通过小鼠逆转录病毒诱变鉴定的基因。长期目标是培养用于生产单克隆抗体的人血浆B细胞系。来自斯坦福大学化学工程系的教师、研究生和本科生将在库比蒂诺高中收养一班新生。这个课程被称为AVID（通过个人决定的进步）。该计划确定学生的课堂表现差，但良好的出勤率和最小的纪律问题。该计划没有基于种族或性别的选择标准，尽管大多数学生在经济上处于不利地位，代表性不足的少数民族将是他们家庭中第一个上大学的人。这个为期四年的项目旨在帮助这些学生从高中毕业并上大学。