Analysis of ABCA Transporter Function in Homologous Recombinant Mice

同源重组小鼠 ABCA 转运蛋白功能分析

• 批准号: 7893990
• 负责人: MASON W FREEMAN
• 金额: $ 44.25万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-26 至 2012-08-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7893990
• 关键词: Analysis; ABCA; Transporter; Function; Homologous

DESCRIPTION (Provided by applicant): The ATP-binding cassette (ABC) transporters compose one of the largest and most important families of transmembrane proteins in the biological kingdom. Within the ABC family is a group of molecules that are referred to as A class transporters that share substantial sequence similarity. The ABCA class molecules comprise twelve proteins of which only two, ABCA1 and ABCA4, have a clearly defined function. This grant proposes to generate homologous recombinant mice lacking each of the other ten A class ABCA transporters and to characterize the resulting functional deficit in the homologous recombinant mice. A novel recombinant bacterial artificial chromosome technology will be used to produce the targeting vectors and fluorescence-in-situ hybridization will be utilized to identify correctly targeted embryonal stem cell clones. Characterization of the null mice will employ high-throughput technologies including liquid chromatography mass spectrometry analysis of tissue lipids and spotted oligonucleotide microarray measurements of gene expression. All of the reagents generated in this work, including null mice, full-length expression cDNAs, and anti-ABCA1 antibodies will be made available to the research community so that other investigators can exploit them in order to facilitate our understanding of the role these ABCA transporters play in human physiology and disease.

描述（由申请人提供）：ATP 结合盒 (ABC) 转运蛋白构成生物界中最大和最重要的跨膜蛋白家族之一。 ABC 家族中有一组被称为 A 类转运蛋白的分子，它们具有显着的序列相似性。 ABCA 类分子包含 12 种蛋白质，其中只有 ABCA1 和 ABCA4 两种蛋白质具有明确定义的功能。该资助计划产生缺乏其他十种 A 类 ABCA 转运蛋白的同源重组小鼠，并表征同源重组小鼠中由此产生的功能缺陷。新型重组细菌人工染色体技术将用于生产靶向载体，并利用荧光原位杂交来识别正确靶向的胚胎干细胞克隆。无效小鼠的表征将采用高通量技术，包括组织脂质的液相色谱质谱分析和基因表达的斑点寡核苷酸微阵列测量。这项工作中产生的所有试剂，包括无效小鼠、全长表达 cDNA 和抗 ABCA1 抗体，都将提供给研究界，以便其他研究人员可以利用它们，以促进我们了解这些 ABCA 的作用转运蛋白在人类生理和疾病中发挥作用。