Transgenic Organ Cultures

转基因器官培养

基本信息

  • 批准号:
    6599686
  • 负责人:
  • 金额:
    $ 8.09万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2003
  • 资助国家:
    美国
  • 起止时间:
    2003-08-06 至 2005-01-30
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): Our knowledge of the molecular basis for kidney development has expanded dramatically over the last 10 years. Much of this new information has come from the study of transgenic and gene-targeted mice. However, there are serious limitations to this experimental approach. First, there is the cost and time involved in the production of each strain of knockout mouse. Secondly, taking an example from our own work with the Wtl gene, the early demise of the metanephric rudiment leaves little actual material to study in order to gain further insight as to the mechanism by which Wt1 is involved in early kidney development [3, 4]. Further study of the Wt1 gene in tissue culture settings has added little to our understanding of its role in early kidney development [5]. Because of these limitations, we have sought to develop alternative and less costly ways of studying the molecular basis of early kidney development. Recently, several studies have demonstrated that it is possible to use electroporation to achieve gene transfer into embryonic tissues; this has found particular use in the study of neural tube and limb development [6-10]. In our laboratory, we are adapting electroporation technology to achieve gene transfer into metanephric kidney organ cultures. In contrast to work with strains of mutant mice, several advantages are immediately apparent. (1) there can be much higher throughput in studying the effects of specific genes on kidney development; (2) wild type organ cultures are used, hence every embryo provides 2 organ cultures, instead of the 25 % of embryos from a litter that might be homozygous for a mutation; (3) in cases like Wt1, where the knockout causes apoptosis, alternate experimental approaches are available, for example those that examine gain of function phenotypes. Microinjection and electroporation will be used to elucidate the role of WT1 in early kidney development. Hypotheses relating to whether Wtl regulates various growth factors will be tested by ectopically expressing or inhibiting Wt1 in the metanephric mesenchyme, and examining the expression of potential target genes by in situ hyridization.
描述(由申请人提供):在过去的10年里,我们对肾脏发育的分子基础知识有了显著的扩展。这些新信息大多来自对转基因和基因靶向小鼠的研究。然而,这种实验方法有严重的局限性。首先,每一株基因敲除小鼠的生产都涉及成本和时间。其次,以我们自己对Wtl基因的研究为例,后肾雏形的早期消亡几乎没有留下实际的材料来研究,以进一步了解Wt1参与早期肾脏发育的机制[3,4]。在组织培养环境下对Wt1基因的进一步研究使我们对其在早期肾脏发育中的作用的理解很少。由于这些限制,我们已经寻求开发替代和更便宜的方法来研究早期肾脏发育的分子基础。最近,一些研究表明,利用电穿孔实现基因转移到胚胎组织是可能的;这在神经管和肢体发育的研究中得到了特别的应用[6-10]。在我们的实验室里,我们正在采用电穿孔技术将基因转移到后肾器官培养中。与使用突变小鼠相比,有几个优点是显而易见的。(1)研究特定基因对肾脏发育的影响可以有更高的通量;(2)使用野生型器官培养,因此每个胚胎提供2个器官培养,而不是一窝胚胎中25%的胚胎可能是纯合的突变;(3)在像Wt1这样的病例中,基因敲除导致细胞凋亡,可采用其他实验方法,例如检查功能表型的获得。显微注射和电穿孔将用于阐明WT1在早期肾脏发育中的作用。关于Wtl是否调节各种生长因子的假设将通过异位表达或抑制后肾间质Wt1来验证,并通过原位杂交检测潜在靶基因的表达。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

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Jordan A Kreidberg其他文献

KIDNEY AND LUNG ABNORMALITIES IN α-3 INTEGRIN DEFICIENT MICE. † 338
α-3 整合素缺陷小鼠的肾脏和肺部异常。†338
  • DOI:
    10.1203/00006450-199604001-00358
  • 发表时间:
    1996-04-01
  • 期刊:
  • 影响因子:
    3.100
  • 作者:
    Stuart L Goldstein;Michael J Donovan;Jordan A Kreidberg
  • 通讯作者:
    Jordan A Kreidberg

Jordan A Kreidberg的其他文献

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{{ truncateString('Jordan A Kreidberg', 18)}}的其他基金

The Role of Fibronectin in ADPKD
纤连蛋白在 ADPKD 中的作用
  • 批准号:
    10217127
  • 财政年份:
    2019
  • 资助金额:
    $ 8.09万
  • 项目类别:
The Role of Fibronectin in ADPKD
纤连蛋白在 ADPKD 中的作用
  • 批准号:
    10021645
  • 财政年份:
    2019
  • 资助金额:
    $ 8.09万
  • 项目类别:
Transcriptional Reprogramming in Podocyte Injury
足细胞损伤中的转录重编程
  • 批准号:
    9149798
  • 财政年份:
    2016
  • 资助金额:
    $ 8.09万
  • 项目类别:
The role of beta-catenin in cyst initiation in Autosomal Dominant Polycystic Kidn
β-连环蛋白在常染色体显性多囊肾囊肿发生中的作用
  • 批准号:
    9064636
  • 财政年份:
    2014
  • 资助金额:
    $ 8.09万
  • 项目类别:
The role of beta-catenin in cyst initiation in Autosomal Dominant Polycystic Kidn
β-连环蛋白在常染色体显性多囊肾囊肿发生中的作用
  • 批准号:
    8683329
  • 财政年份:
    2014
  • 资助金额:
    $ 8.09万
  • 项目类别:
Misregulation of receptor tyrosine kinase signaling in PKD
PKD 中受体酪氨酸激酶信号传导的失调
  • 批准号:
    8338906
  • 财政年份:
    2011
  • 资助金额:
    $ 8.09万
  • 项目类别:
Misregulation of receptor tyrosine kinase signaling in PKD
PKD 中受体酪氨酸激酶信号传导的失调
  • 批准号:
    8726973
  • 财政年份:
    2011
  • 资助金额:
    $ 8.09万
  • 项目类别:
Misregulation of receptor tyrosine kinase signaling in PKD
PKD 中受体酪氨酸激酶信号传导的失调
  • 批准号:
    8238482
  • 财政年份:
    2011
  • 资助金额:
    $ 8.09万
  • 项目类别:
Misregulation of receptor tyrosine kinase signaling in PKD
PKD 中受体酪氨酸激酶信号传导的失调
  • 批准号:
    8541009
  • 财政年份:
    2011
  • 资助金额:
    $ 8.09万
  • 项目类别:
BMP and FGF Signaling in kidney progenitor cells
肾祖细胞中的 BMP 和 FGF 信号转导
  • 批准号:
    8494042
  • 财政年份:
    2010
  • 资助金额:
    $ 8.09万
  • 项目类别:

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