Analysis of ABCA Transporter Function in Homologous Recombinant Mice

描述(申请人提供)：三磷酸腺苷结合盒(ABC)转运体是生物界中最大和最重要的跨膜蛋白家族之一。在ABC家族中，有一组分子被称为A类转运蛋白，它们有很大的序列相似性。ABCA类分子由12种蛋白质组成，其中只有两种具有明确定义的功能：ABCA1和ABCA4。这项资助建议培育缺乏其他10个A类ABCA转运蛋白的同源重组小鼠，并鉴定由此产生的同源重组小鼠的功能缺陷。将使用一种新的重组细菌人工染色体技术来生产靶向载体，并将利用荧光原位杂交来正确识别靶向胚胎干细胞克隆。对缺失小鼠的鉴定将使用高通量技术，包括组织脂类的液-质联用分析和基因表达的斑点寡核苷酸微阵列测量。在这项工作中产生的所有试剂，包括空小鼠、全长表达的cDNA和抗ABCA1抗体将提供给研究界，以便其他研究人员可以利用它们，以促进我们了解这些ABCA转运蛋白在人类生理和疾病中所起的作用。

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.