NON-TUMORIGENIC TERATOCARCINOMA

非致瘤性畸胎癌

• 批准号: 2093291
• 负责人: PAULETTE J MCCORMICK
• 金额: $ 19.61万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2093291
• 关键词: Retroviridae; cell differentiation; complementary DNA; gene expression; genetic transcription; genetically modified animals; genome; human fetus tissue; laboratory mouse; molecular cloning; neoplastic cell; nucleic acid sequence; open reading frames; phenotype; regulatory gene; teratoma; transfection; transposon /insertion element; viral carcinogenesis

It has become increasingly clear that certain genes, depending upon the time and nature of their expression, can regulate the somewhat opposed processes of both tumorigenesis and differentiation. The embryonal carcinoma (EC) cell system has proven to be of particular value in studies of these two processes as well as in evaluating their interrelatedness. Therefore, in an attempt to identify novel regulatory genes the applicant mutagenized an EC cell line via retroviral insertion and generated a mutant line that differs from EC cells in both tumorigenicity and differentiation. The applicant has shown that the mutation was created by a single insertion into a hemizygous site. Sequencing of both the immediate 5' and 3' flanking regions at this site has revealed open reading frames with no homology to any known mouse gene. This indicates that the mutant phenotypes may have arisen as a result of viral insertion into a novel mouse coding sequence. The applicant proposes to use further flanking sequence information, zoo blotting, CpG island analysis and exon trapping techniques to identify a transcription unit at or near this locus. This transcript will be used to isolate a full length cDNA clone and then compare and sequence the gene from both cDNA and genomic clones and examine the 5' regulatory region. The applicant will also analyze the functional consequences of expression of the transcript in cell transfectants and transgenic mice and determine the tissue and embryonic specific expression patterns. Finally, she will isolate and characterize specific proteins whose expression has been altered as a consequence of the gene disruption. The applicant believes that these studies will result in the identification of a novel mouse gene and contribute to our understanding of the molecular and cellular elements controlling both proliferation and differentiation.

越来越清楚的是，某些基因，取决于 时间和性质，他们的表达，可以调节有点反对 肿瘤发生和分化的过程。 胚胎 癌（EC）细胞系统已被证明在研究中具有特别的价值 这两个过程，以及在评估它们的相互关系。 因此，为了鉴定新型调节基因，申请人 通过逆转录病毒插入诱变EC细胞系，并产生 在致瘤性和致突变性方面均不同于EC细胞的突变系 分化 申请人已证明突变是由 在半合子的位置上插入一个细胞 两个序列的测序 在该位点的直接5'和3'侧翼区揭示了开放的 阅读框与任何已知的小鼠基因都没有同源性。这表明 突变表型可能是病毒插入的结果 一个新的小鼠编码序列。 申请人建议进一步使用 侧翼序列信息，动物园印迹，CpG岛分析和外显子 捕获技术，以识别在该位置或附近的转录单位， 基因座 该转录本将用于分离全长cDNA克隆 然后将cDNA和基因组克隆的基因进行比较和测序， 并检查5'调控区。 申请人还将分析 转录本在细胞中表达的功能后果 转染子和转基因小鼠，并确定组织和胚胎 特定的表达模式。 最后，她会分离并描述 特定蛋白质，其表达已被改变，作为结果， 基因破坏 申请人认为，这些研究将 结果发现了一个新的小鼠基因，并有助于我们 了解控制两者的分子和细胞元素 增殖和分化。