Tuberous sclerosis complex (TSC) is a rare autosomal dominant disorder involving multiple organ systems. TSC2 gene plays an important role in the development of TSC. The most common kidney manifestation of TSC is renal angiomyolipoma (RAML). TSC-RAML is more likely to be bilateral multiple tumors and tends to destroy the renal structure and damages renal function severely. As a result, patients with TSC-RAML often miss the opportunity for surgical treatment when TSC-RAML is diagnosed, causing difficulty in obtaining tumor specimens through surgery. Due to this difficulty, model cell lines must be constructed for scientific research. In this paper, TSC2 was knocked out in NIH-3T3 cell lines by CRISPR/Cas9 system. PCR, WB and mTOR inhibitor drug sensitivity test showed that the TSC2 knockout NIH-3T3 cells were successfully constructed. The ability of proliferation and invasion in TSC2 KO NIH-3T3 cells were higher than those in wild type group. The constructed KO cell line lay the foundation for further study of TSC.

结节性硬化症 (TSC) 是一种罕见的常染色体显性遗传病，涉及多个器官系统。TSC2基因在TSC的发生发展中起重要作用。TSC 最常见的肾脏表现是肾血管平滑肌脂肪瘤 (RAML)。TSC-RAML更可能为双侧多发性肿瘤，并有破坏肾脏结构和严重损害肾功能的倾向。因此，TSC-RAML患者往往在诊断为TSC-RAML时错过了手术治疗的机会，导致难以通过手术获得肿瘤标本。由于这一困难，必须构建模型细胞系用于科学研究。本文利用CRISPR/Cas9系统在NIH-3T3细胞系中敲除TSC2。PCR、WB和mTOR抑制剂药敏试验表明，成功构建了TSC2基因敲除的NIH-3T3细胞。TSC2 KO NIH-3T3细胞的增殖和侵袭能力高于野生型组。构建的KO细胞系为进一步研究TSC奠定了基础。