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文献推荐 | CRISPR/Cas9技术敲除斑马鱼rpl15基因对斑马鱼红系造血发育的影响
2023-10-23
[摘要] 目的 利用CRISPR/Cas9技术敲除斑马鱼rpl15基因，探讨rpl15基因对斑马鱼红系造血发育的影响。方法 针对斑马鱼rpl15基因设计并制备gRNA(guide RNA)，将gRNA与Cas9蛋白的混合物显微注射到单细胞期斑马鱼受精卵中，提取72 h胚胎基因组DNA，PCR扩增出目的片段，T7E1限制性酶切法检测基因打靶情况，随后进行测序分析确定编辑效果。通过O-dianisidine染色分析血红蛋白合成情况，中性粒细胞染色分析中性粒细胞生成情况，利用Real-time PCR检测p53、mdm2、hsp70和造血相关转录因子基因的表达变化。结果 成功制备了rpl15 gRN......
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