Abstract: Objective The CSF-1R knock in mouse model was established by CRISPER / Cas9 technology. Method The partial sequences of exons 3 to 11 cDNA of humanized CSF-1R gene, together with TGA codon and WPREPolyA original, were inserted into downstream of the 20th alanine of exon 3 on mouse CSF-1R gene. The gene expression characteristics were further analyzed by RT-qPCR and sperm cryopreservation was carried out for transgene positive mice. Result The result showed that the trans gene expressed in heart, liver, spleen, lung, kidney and brain of CSF-1R gene knock-in mice. The average in vitro fertilization rate of frozen sperm was 67. 74%, and the average birth rate was 16. 06%. Conclusion A humanized CSF-1R gene knock-in mice model was established successfully,which provides a novel tool for evaluating the in vivo efficacy of antibodies and drugs of this target.

Key words: CRISPR-Cas9, CSF-1R gene, humanized mouse model, antibody drugs

摘要： 目的 利用 CRISPR-Cas9 技术构建 CSF-1R 基因敲入小鼠模型。 方法 将人 CSF-1R 基因的 3 到 11 外显子 cDNA 的部分序列,加上 TGA 密码子及 WPRE-PolyA 原件,插入到了小鼠 CSF-1R 基因外显子 3 的第 20 位丙氨酸下游。 在鉴定获得阳性模型后,进一步通过 RT-qPCR 分析基因的表达特征并进行精子冷冻保种繁殖。 结果 该基因正确插入设计位点,转录水平显示转入基因能在小鼠模型心脏、肝、脾、肺、肾、脑等脏器中表达,冷冻精子复苏后体外受精的平均受精率达 67. 74% ,平均产仔率 16. 06% ,并经过 PCR 检测得到阳性小鼠。 结论 成功构建了人源化CSF-1R 基因敲入小鼠模型,为评价肿瘤免疫抗体药效及相关靶向抗肿瘤药物筛选提供了动物模型。

关键词: CRISPR / Cas9 技术, CSF-1R 基因, 人源化小鼠模型, 抗体药物