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RETROTRANSPOSITION IN L1 TRANSGENIC MICE

L1 转基因小鼠的逆转录转座

基本信息

批准号：
6748147
负责人：
ELINE Tjetske LUNING PRAK
金额：
$ 13.66万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-04-10 至 2005-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6748147
关键词：
HeLa cells Retroviridae beta galactosidase genetic markers genetic transcription genetically modified animals green fluorescent proteins laboratory mouse mutagens nucleic acid sequence plasmids transfection transfection /expression vector transposon /insertion element

项目摘要

L1 transposons are mobile DNA elements which are known to be active in several species including mouse and man. Their activity is known to cause disease. L1 retrotransposons move via a copy and paste mechanism and exhibit no known market integration site preferences. These features make them useful as insertional mutagens because they can tag the genes they disrupt. A insertional mutagens, L1 retrotransposons could be used to study tumor progression. Through genetic engineering they potentially could be designed to move in a tissue-specific, developmental stage- specific fashion. As such, they could revolutionize cancer genetics. However, the activity of L1 transposons in vivo is poorly understood. The frequency with which they move about is not known. It also is not well understood where and when retrotransposons are active. This proposal is designed to further our understanding of the biology of L1 retrotransposons in vivo and establish whether they can serve as insertional mutagens. To achieve these aims we will demonstrate L1 retrotransposition from integrated chromosomal sites. Next we will create a screenable marker assay for retrotransposition. This screenable marker assay will allow us to characterize the frequency of retrotransposition events without prolonged selections in antibiotics. We will use this assay to evaluate the cell types and states in which retrotransposition events are favored. We hope to address whether retrotransposition events are favored. We hope to address whether retrotransposition events occur more frequently in the transformed state and promote genetic instability in tumor cells. Finally, we will establish an in vivo model of L1 retrotransposition by adapting our L1 reporter constructs to maximize the chances of in vivo detection. We will breed our L1 transgenics to ubiquitously expressing lacZ indicator mice. Mice containing both the tagged L1 retrotransposon and the screenable marker allele will be used to eliminate the frequency of retrotransposition in vivo and determine the tissue types and developmental stages in which retrotransposition events occur.

L1 transposons are mobile DNA elements which are known to be active in several species including mouse and man.已知它们的活动会导致疾病。 L1 retrotransposons move via a copy and paste mechanism and exhibit no known market integration site preferences. These features make them useful as insertional mutagens because they can tag the genes they disrupt. A insertional mutagens, L1 retrotransposons could be used to study tumor progression. Through genetic engineering they potentially could be designed to move in a tissue-specific, developmental stage- specific fashion. As such, they could revolutionize cancer genetics. However, the activity of L1 transposons in vivo is poorly understood. The frequency with which they move about is not known. It also is not well understood where and when retrotransposons are active. This proposal is designed to further our understanding of the biology of L1 retrotransposons in vivo and establish whether they can serve as insertional mutagens.为了实现这些目标，我们将演示来自整合染色体位点的 L1 逆转录转座。接下来，我们将创建用于逆转录转座的可筛选标记测定。 This screenable marker assay will allow us to characterize the frequency of retrotransposition events without prolonged selections in antibiotics.我们将使用此测定来评估有利于逆转录转座事件的细胞类型和状态。我们希望解决逆转录转座事件是否受到青睐的问题。 We hope to address whether retrotransposition events occur more frequently in the transformed state and promote genetic instability in tumor cells. Finally, we will establish an in vivo model of L1 retrotransposition by adapting our L1 reporter constructs to maximize the chances of in vivo detection.我们将用 L1 转基因小鼠培育普遍表达 lacZ 的指示小鼠。 Mice containing both the tagged L1 retrotransposon and the screenable marker allele will be used to eliminate the frequency of retrotransposition in vivo and determine the tissue types and developmental stages in which retrotransposition events occur.