The Knockout of the Mouse Muc5ac

小鼠 Muc5ac 的敲除

• 批准号: 7594029
• 负责人: Robert Langenbach
• 金额: $ 45.76万
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7594029
• 关键词: Cells; Chimera organism; Gastric ulcer; Genes; Genomics; Human; Infection; Inflammation; Knockout Mice; Lung; Molecular; Mucins; Mus; Neomycin; PTGS2 gene; Pathway interactions; Pharmacology; Physiological; Polymerase Chain Reaction; Predisposition; Reporting; Role; Stomach; blastocyst; embryonic stem cell; insight; malignant stomach neoplasm; transmission process

A targeting construct containing the neomycin and HSTK genes and 10 kb of genomic homology has been constructed to disrupt the Muc5ac gene. The construct has been electroporated into mouse ES cells but to date only 2 PCR positive clones of about 1,000 analyzed have been obtained. These clones are being further analyzed by PCR and genomic Southern analysis to verify targeting. Once targeting is confirmed these clones will be injected into mouse blastocyst to generate chimeras and achieve germline transmission.

我们构建了一个包含新霉素和HSTK基因以及10kb基因组同源性的靶向构建体来破坏Muc5ac基因。该结构已被电穿孔到小鼠胚胎干细胞中，但迄今为止，仅获得了约1000个PCR阳性克隆。这些克隆正在通过PCR和基因组Southern分析进一步分析，以验证靶向性。一旦确定靶向性，这些克隆将被注射到小鼠囊胚中产生嵌合体并实现种系传播。