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文獻推薦 | CRISPR/Cas9技術(shù)敲除斑馬魚rpl15基因?qū)Π唏R魚紅系造血發(fā)育的影響
2023-10-23
[摘要] 目的 利用CRISPR/Cas9技術(shù)敲除斑馬魚rpl15基因肌索，探討rpl15基因?qū)Π唏R魚紅系造血發(fā)育的影響恋谭。方法 針對斑馬魚rpl15基因設(shè)計并制備gRNA(guide RNA)弥虐，將gRNA與Cas9蛋白的混合物顯微注射到單細胞期斑馬魚受精卵中，提取72 h胚胎基因組DNA脏毯，PCR擴增出目的片段闹究，T7E1限制性酶切法檢測基因打靶情況，隨后進行測序分析確定編輯效果食店。通過O-dianisidine染色分析血紅蛋白合成情況渣淤，中性粒細胞染色分析中性粒細胞生成情況赏寇，利用Real-time PCR檢測p53、mdm2价认、hsp70和造血相關(guān)轉(zhuǎn)錄因子基因的表達變化嗅定。結(jié)果 成功制備了rpl15 gRN......
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