NON TUMORIGENIC TERATOCARCINOMA CELL LINE
非致瘤性畸胎癌细胞系
基本信息
- 批准号:6137472
- 负责人:
- 金额:$ 23.99万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1990
- 资助国家:美国
- 起止时间:1990-07-01 至 2001-03-31
- 项目状态:已结题
- 来源:
- 关键词:RNase protection assay Retroviridae athymic mouse cell differentiation complementary DNA embryonic stem cell gene expression gene mutation gene targeting genetic library genetic regulation genetic regulatory element genetically modified animals messenger RNA mutant northern blottings nucleic acid hybridization nucleic acid sequence phenotype polymerase chain reaction teratoma tissue /cell culture transfection transposon /insertion element viral carcinogenesis
项目摘要
Embryonal carcinoma (EC) cells have proven to be of tremendous value in
analyses of both oncogenesis and mammalian development as well as in
studies of the interrelatedness of these two processes. Of particular note
are the use of these cells in the development of transgenic animals and in
studies of retinoid mediated differentiation therapy. Previously we
infected an EC cell line (NR1-0) with a defective retrovirus containing a
neomycin resistance cassette (NeoR) and created a mutant cell line
designated NR1-6. This cell line is unique in its morphological, adhesive,
tumorigenic and differentiative properties. Genetic analyses of mutant,
hybrid and revertant cell lines indicate that there is only a single
retroviral insertion site regulating all of these recessive, and,
apparently, genetically linked, phenotypess. To elucidate the molecular
regulation of this pleiotropic mutation we have sequenced 17.1 kb of
genomic DNA flanking the insertion site. With the exception of repeat
elements, this region shares no significant homology with any previously
reported sequence and is therefore a novel locus. Using a variety of
techniques we searched for transcripts encoded both within this region and
elsewhere in the genome whose expression might be directly or indirectly
regulated by the insertion site locus.
We have identified at least three novel exons encoded within the 17 kb
flanking sequence, two of which hybridize to a transcript of 3.3 kb in
northern analysis of mouse cells and tissues. Using differential display
and mRNA fingerprinting we have also identified a number of potential
downstream target genes whose expression differ between the NR1-6 and NR1-0
cell lines. We now propose to evaluate these findings in more detail in
order to understand the molecular mechanisms regulating the original NR1-6
mutation and to identify the downstream acting loci which regulate the
variant phenotypes (most particularly differentiative response to
retinoids) associated with the NR1-6 mutation. To this end we will conduct
further molecular and functional analyses of: the transcript(s) encoded by
the insertion locus (Specific Aim 1), and transcripts encoded by other loci
which have been shown to be differentially expressed between parent and
mutant cells (Specific Aim2).
胚胎癌(EC)细胞已被证明具有巨大的价值
分析肿瘤发生和哺乳动物发育以及
研究这两个过程的相互关系。 特别值得注意的是
这些细胞在转基因动物的发育中的用途以及
类维生素A介导的分化治疗的研究。 以前我们
用含有缺陷的逆转录病毒感染 EC 细胞系 (NR1-0)
新霉素抗性盒(NeoR)并创建了突变细胞系
指定为NR1-6。 该细胞系在形态、粘附性、
致瘤和分化特性。 突变体的遗传分析,
杂交和回复细胞系表明只有一个
逆转录病毒插入位点调节所有这些隐性,并且,
显然,遗传相关,表型。 为了阐明分子
我们已经对这种多效性突变的调节进行了 17.1 kb 的测序
插入位点两侧的基因组 DNA。 除了重复之外
元素,该区域与之前的任何区域没有显着的同源性
报道的序列,因此是一个新的基因座。 使用各种
我们搜索该区域内编码的转录本的技术
基因组中其他地方的表达可能直接或间接
受插入位点基因座调节。
我们已经鉴定出至少三个编码在 17 kb 内的新外显子
侧翼序列,其中两个与 3.3 kb 的转录物杂交
小鼠细胞和组织的 Northern 分析。 使用差分显示
和 mRNA 指纹识别,我们还发现了一些潜在的
NR1-6 和 NR1-0 之间表达不同的下游靶基因
细胞系。 我们现在建议更详细地评估这些发现
为了了解调节原始NR1-6的分子机制
突变并鉴定调节下游作用位点
变异表型(最特别的是对
类视黄醇)与 NR1-6 突变相关。 为此我们将进行
进一步的分子和功能分析: 编码的转录物
插入基因座(具体目标 1),以及其他基因座编码的转录本
已被证明在亲本和亲本之间表达存在差异
突变细胞(具体目标2)。
项目成果
期刊论文数量(0)
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科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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PAULETTE J MCCORMICK其他文献
PAULETTE J MCCORMICK的其他文献
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{{ truncateString('PAULETTE J MCCORMICK', 18)}}的其他基金
Analyses of a Non tumorigenic Teratocarcinoma Cell Line
非致瘤性畸胎癌细胞系的分析
- 批准号:
6512638 - 财政年份:1990
- 资助金额:
$ 23.99万 - 项目类别:
ANALYSIS OF A NON-TUMORIGENIC TERATOCARCINOMA CELL LINE
非致瘤性畸胎癌细胞系的分析
- 批准号:
3193573 - 财政年份:1990
- 资助金额:
$ 23.99万 - 项目类别:
ANALYSIS OF A NON-TUMORIGENIC TERATOCARCINOMA CELL LINE
非致瘤性畸胎癌细胞系的分析
- 批准号:
3193577 - 财政年份:1990
- 资助金额:
$ 23.99万 - 项目类别: