Vector Integration and Tracking

矢量积分和跟踪

基本信息

  • 批准号:
    8567347
  • 负责人:
  • 金额:
    $ 43.46万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
  • 资助国家:
    美国
  • 起止时间:
  • 项目状态:
    未结题

项目摘要

The development of leukemia has been observed in 5 out of 19 SCID-Xl patients treated by stem cell gene therapy. In all 5 patients the development of leukemia was due to insertional mutagenesis, i.e. the retrovirus- mediated activation of nearby proto-oncogenes. Thus, the careful study of integration sites and the contribution of individual clones to repopulation will be of crucial importance for all gene therapy studies, and the FDA has mandated the careful monitoring of retroviral integration sites in all clinical gene therapy studies. The Vector Integration and Tracking Core D will provide all projects a centralized facility to efficiently identify foamy virus (FV) vector integration sites in complex biological DNA samples from in vitro or in vivo studies. We will use recently developed improved non-restriction (nr)LAM-PCR. The core will be able to process a variety of samples from murine, dog, or human to generate DNA for shuttle vector or PCR- based methods. For nrLAM-PCR, as little as lOng of DNA can be used to carry out FV vector integration site amplification and extended processing in preparation for sequencing. The samples processed by Core D will be compatible with both Sanger-based sequencing for pilot experiments and new sequencing methodologies, such as pyrosequencing, for deep sequencing to identify all of the amplifiable FV vector integration sites in a given sample. Core D will also provide assistance to all projects to design primers specific to FV vector integration sites to allow for DNA-based real time (RT)-PCR tracking to assess the contribution for individual clones that are deemed important for further investigation. In addition, Core D will provide a centralized facility to analyze integration sites via a common gateway interface (CGI)-PERL web server. Current versions of the human (hg19), dog (canFam2) and mouse (mm9) genomes will be supported. The Core will also provide support to investigators through PERL programs to correlate FV vector integration sites with data from published databases including proto-oncogene TSS, microarray data and over- represented gene classes. The bioinformatics component will also generate random datasets for all three genomes, human, mouse and dog, to evaluate over-represented gene classes near vector proviruses.
在19例接受干细胞基因治疗的SCID-X1患者中,有5例发生白血病。在所有5名患者中,白血病的发生是由于插入诱变,即逆转录病毒介导的附近原癌基因的激活。因此,对整合位点的仔细研究和单个克隆对再增殖的贡献对于所有基因治疗研究都至关重要,并且FDA已授权在所有临床基因治疗研究中仔细监测逆转录病毒整合位点。载体整合和跟踪核心D将为所有项目提供一个集中的设施,以有效地识别体外或体内研究中复杂生物DNA样品中的泡沫病毒(FV)载体整合位点。我们将使用最近开发的改进的非限制性(nr)LAM-PCR。核心将能够处理来自鼠、狗或人的各种样品,以产生用于穿梭载体或基于PCR的方法的DNA。对于nrLAM-PCR,可以使用少至10 ng的DNA来进行FV载体整合位点扩增和延伸加工以准备测序。由核心D处理的样品将与用于中试实验的基于Sanger的测序和用于深度测序的新测序方法(如焦磷酸测序)兼容,以鉴定给定样品中的所有可扩增的FV载体整合位点。核心D还将为所有项目提供援助,以设计针对FV载体整合位点的特异性引物,从而进行基于DNA的真实的时间(RT)-PCR跟踪,以评估被认为对进一步研究重要的单个克隆的贡献。此外,Core D将提供一个集中的设施,通过公共网关接口(CGI)-PERL Web服务器来分析集成站点。将支持人类(hg 19)、犬(canFam 2)和小鼠(mm 9)基因组的当前版本。核心还将通过PERL程序为研究者提供支持,以将FV载体整合位点与来自已发表数据库的数据相关联,包括原癌基因TSS、微阵列数据和过度代表的基因类别。生物信息学部分还将为所有三个基因组(人类、小鼠和狗)生成随机数据集,以评估载体前病毒附近过度代表的基因类别。

项目成果

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BRIAN C BEARD其他文献

BRIAN C BEARD的其他文献

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{{ truncateString('BRIAN C BEARD', 18)}}的其他基金

Vector Integration and Tracking
矢量积分和跟踪
  • 批准号:
    8278891
  • 财政年份:
    2012
  • 资助金额:
    $ 43.46万
  • 项目类别:
Development and Safety Evaluation of Chemo-Protective Gene Therapy Vectors
化学保护性基因治疗载体的开发和安全性评价
  • 批准号:
    8136048
  • 财政年份:
    2007
  • 资助金额:
    $ 43.46万
  • 项目类别:
Development and Safety Evaluation of Chemo-Protective Gene Therapy Vectors
化学保护性基因治疗载体的开发和安全性评价
  • 批准号:
    7318585
  • 财政年份:
    2007
  • 资助金额:
    $ 43.46万
  • 项目类别:
Development and Safety Evaluation of Chemo-Protective Gene Therapy Vectors
化学保护性基因治疗载体的开发和安全性评价
  • 批准号:
    7497056
  • 财政年份:
    2007
  • 资助金额:
    $ 43.46万
  • 项目类别:
Development and Safety Evaluation of Chemo-Protective Gene Therapy Vectors
化学保护性基因治疗载体的开发和安全性评价
  • 批准号:
    7681219
  • 财政年份:
    2007
  • 资助金额:
    $ 43.46万
  • 项目类别:
Development and Safety Evaluation of Chemo-Protective Gene Therapy Vectors
化学保护性基因治疗载体的开发和安全性评价
  • 批准号:
    7905203
  • 财政年份:
    2007
  • 资助金额:
    $ 43.46万
  • 项目类别:
Vector Integration and Tracking
矢量积分和跟踪
  • 批准号:
    8712355
  • 财政年份:
  • 资助金额:
    $ 43.46万
  • 项目类别:
Vector Integration and Tracking
矢量积分和跟踪
  • 批准号:
    9117270
  • 财政年份:
  • 资助金额:
    $ 43.46万
  • 项目类别:
Vector Integration and Tracking
矢量积分和跟踪
  • 批准号:
    8897250
  • 财政年份:
  • 资助金额:
    $ 43.46万
  • 项目类别:

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