Generation of a Light Inducible Cre Transgenic Animal for KidneyResearch

摘要 疾病状态的动物模型和用于细胞动力学的原位分析的生物记者是一个重要的方面 肾脏、泌尿学和造血研究；然而，大多数研究集中在 整个生物体或肾脏特定的细胞类型，不能准确反映通常是 与单个细胞或一组细胞中的缺陷有关。这种差异在很大程度上是由于技术上的差异- 能够不受限制地在光学选择的一组细胞中诱导Cre重组的长度 完整的有机体。因此，我们认为，控制基因操作的系统(导致损失 功能等位基因或外源cDNA/生物记者的表达)在活着的有机体的一组细胞中 必填项。该系统不应改变或扰乱周围环境，并应允许潜在的 基于系统的分析和生物标记物的结合。因此，我们假设转基因株系普遍存在于 表达光诱导的Cre将允许这种不可见的光学空间和时间控制 活着的有机体。这种光诱导重组(LIR)系统将在两只小鼠体内建立 以及斑马鱼的模式生物。在完成这项提议后，我们将建立多功能和 用于选择性基因改变的高效工具，将促进区域和潜在的单细胞分析 众多的研究领域。我们预计，这项研究将对许多领域产生广泛的积极影响 肾脏、泌尿学和造血学研究。

ABSTRACT Animal modeling of disease states and bioreporters for in situ analysis of cellular dynamics are an essential aspect of kidney, urological and hematopoietic research; however, most studies focus on genetic manipulation within the whole organism or kidney specific cell types, which does not accurately reflect disease states that are often associated with defects in individual cells or groups of cells. This discrepancy is largely due to the technical chal- lenges of being able to unrestrictedly induce Cre recombination in an optically selected group of cells within an intact organism. Therefore, we believe that a system for controlled genetic manipulation (resulting in loss of function alleles or expression of exogenous cDNAs/bioreporters) within a group of cells of a viable organism is required. This system should not alter or perturb the surrounding environment and should allow for potential systems based analysis and biomarker incorporation. Therefore, we hypothesize that transgenic lines ubiqui- tously expressing a light inducible Cre would allow for such unseen optical spatial and temporal control within a living organism. This Light Inducible Recombination (LIR) systems will be established within both the mouse and the zebrafish model organisms. Upon completion of this proposal we will have established versatile and efficient tools for selective genetic alterations that will facilitate regional and potentially single cell analysis in a multitude of research fields. We anticipate that this research will have a broad positive impact on a number areas of kidney, urological and hematopoietic research.