HOMOLOGOUS RECOMBINATION TO INACTIVATE MOUSE ANP

同源重组使小鼠 ANP 失活

• 批准号: 2133890
• 负责人: MARTIN R. POLLAK
• 金额: $ 7.56万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2133890
• 关键词: atrial natriuretic peptide; blood pressure; blood volume; embryonic stem cell; gene induction /repression; gene mutation; genetic recombination; genetically modified animals; hormone receptor; hormone regulation /control mechanism; inbreeding; laboratory mouse; microinjections; tissue /cell culture; transfection

Atrial natriuretic peptide (ANP) is a recently discovered hormone with diverse physiologic properties. Studies are elucidating the role of ANP in multiple pathologic conditions, but its exact importance remains unclear. The gene for mouse ANP has been cloned and sequenced in the sponsor's laboratory. The goal of the Phase I project will be to inactivate the ANP gene in mice. Specifically, the aims are to 1) Construct embryonic stem (ES) cells with a defective ANP gene by homologous recombination; 2) Microinject ES cells with the mutant ANP gene into mouse blastocysts; 3) Transfer these blastocysts into the uteri of pseudopregnant mice; 4) Determine which of the resulting chimeric mice have the mutant ANP gene in the germ line; 5) Interbreed these chimeras to produce mice homo- and heterozygous for the ANP gene deletion. Physiologic consequences of ANP deletion will then be assessed. It is expected that the heterozygotes will be viable; the viability of the homozygotes is less certain. If not viable, the developing ANP-/ANP- embryos will be studied histologically to define the effects of ANP deletion. During Phase II of this grant, similar methods will be used to further characterize the physiology of ANP by utilizing homologous recombination to inactivate the ANP receptor genes. These studies will aid in the investigation of ANP function in normal and pathologic states. This may prove to be of particular import in helping to understand the dysregulation of blood pressure and volume homeostasis seen in liver failure, heart disease, renal disease, and hypertension.

心钠素(ANP)是一种新近发现的激素。 不同的生理特性。研究正在阐明心钠素的作用 在多种病理条件下，但其确切的重要性仍然存在 不清楚。小鼠心钠素基因已被克隆和测序 赞助商的实验室。第一阶段项目的目标将是 使小鼠的心钠素基因失活。具体地说，目标是1) 利用ANP缺陷基因构建胚胎干细胞 同源重组；2)用突变型ANP显微注射ES细胞 将基因导入小鼠胚泡；3)将这些胚泡移植到子宫 假孕小鼠；4)确定产生的嵌合体小鼠中的哪一个 有突变的ANP基因在胚系中；5)使这些嵌合体杂交 产生ANP基因缺失的纯合子和杂合子小鼠。 然后将评估ANP缺失的生理后果。它是 预计杂合子将是可行的； 纯合子就不那么确定了。如果不可行，正在开发的ANP-/ANP- 将对胚胎进行组织学研究，以确定ANP的作用 删除。在这笔赠款的第二阶段，将使用类似的方法来 利用同源基因进一步研究ANP的生理学特征 重组使心钠素受体基因失活。这些研究将 协助研究ANP在正常和病理状态下的功能。 这可能会被证明对帮助理解 肝脏出现的血压和容量平衡失调 衰竭、心脏病、肾脏疾病和高血压。