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发明授权
CN106754917B 胶质母细胞瘤中同时敲除EGFRwt和EGFRvIII的方法 有权

胶质母细胞瘤中同时敲除EGFRwt和EGFRvIII的方法
摘要：
本发明公开了一种胶质母细胞瘤中同时敲除EGFRwt和EGFRvIII的方法，涉及DNA重组技术。本发明中的sgRNA在EGFR基因的靶向位点位于EGFR基因的第17号外显子上，是表达EGFR跨膜区的基因序列。本发明通过分别构建Cas9和sgRNA慢病毒，并将二者感染胶质瘤细胞的方法，从而达到同时敲除胶质瘤细胞中EGFRwt/vIII的目的。使用本发明所述方法可同时稳定的敲除EGFR wt和EGFRvIII两个基因，敲除后可有效的抑制肿瘤发生发展。本发明所述sgRNA有望在治疗EGFR扩增和EGFRvIII突变型胶质母细胞瘤的新型药物中得到应用。
摘要（英）：

The invention discloses a method for knocking out EGFRwt and EGFRvIII simultaneously from glioblastoma multiforme and relates to the recombinant DNA technology. The target site of sgRNA in an EGFR gene is on exon 17 of the EGFR gene, and is a gene sequence expressing a transmembrane region of the EGFR. According to the method for constructing Cas9 and sgRNA lentiviruses and infecting the glioblastoma multiforme with the lentiviruses, EGFRwt and EGFRvIII can be knocked out simultaneously from the glioblastoma multiforme. EGFRwt and EGFRvIII can be knocked out simultaneously and stably from the glioblastoma multiforme with the method, and occurrence and development of tumor can be inhibited effectively after knockout. The sgRNA is expected to be applied to novel drugs for treating EGFR amplification type and EGFRvIII mutant type glioblastoma multiforme.

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