Anaplasia & differentiation in a reversible cancer model

发育不全

• 批准号: 6585101
• 负责人: DANIEL J MURPHY
• 金额: $ 4.81万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-11 至 2006-06-10
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6585101
• 关键词: 3T3 cells; apoptosis; carcinogenesis; cell differentiation; cell growth regulation; cytogenetics; dissection; flow cytometry; gene induction /repression; gene interaction; genetically modified animals; immunocytochemistry; keratinocyte; laboratory mouse; microarray technology; neoplasm /cancer genetics; postdoctoral investigator; protooncogene; skin neoplasms; transfection; tumor suppressor genes

DESCRIPTION (provided by applicant): A reversible transgenic mouse model for skin cancer will be used to investigate the relationship between neoplasia and anaplasia, with the long term goal of better understanding the consequences of oncogene activation and inactivation for differentiation. The specific aims are 1) to test the hypothesis that neoplastic INV-MycERTM keratinocytes will differentiate upon removal of ectopic Myc function; 2) to identify the in vivo alterations in keratinocyte gene expression upon activation and subsequent deactivation of c-MycERTM; 3) to engineer a ubiquitously prospective, conditionally expressed, regulatable c-Myc transgenic mouse. IHC and ISH will be employed to assess the differentiation state of INV-MycERTM keratinocytes in SA1 while tissue microdissection coupled with high throughput microarray analysis will define the transcriptional basis for Myc-induced change in SA2. For SA3, the MycERTM transgene, linked by way of an IRES to EGFP and preceded by a floxed Stop cassette, will be targeted to a ubiquitously active locus by homologous recombination.

描述（由申请人提供）：一个可逆的转基因皮肤癌小鼠模型将用于研究肿瘤和非增生之间的关系，其长期目标是更好地了解癌基因激活和失活对分化的影响。具体目的是：1)验证肿瘤INV-MycERTM角质形成细胞在去除异位Myc功能后会分化的假设；2)鉴定c-MycERTM激活和失活后角质形成细胞基因在体内的表达变化；3)设计一种具有普遍前景的、有条件表达的、可调节的c-Myc转基因小鼠。IHC和ISH将用于评估SA1中INV-MycERTM角质形成细胞的分化状态，而组织显微解剖结合高通量微阵列分析将确定myc诱导SA2变化的转录基础。对于SA3， MycERTM转基因，通过IRES连接到EGFP，前面有一个固定的Stop盒，将通过同源重组靶向到一个无处不在的活性位点。