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文獻(xiàn)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，探討rpl15基因?qū)Π唏R魚紅系造血發(fā)育的影響土思。方法 針對斑馬魚rpl15基因設(shè)計(jì)并制備gRNA(guide RNA)玛迄，將gRNA與Cas9蛋白的混合物顯微注射到單細(xì)胞期斑馬魚受精卵中年局，提取72 h胚胎基因組DNA，PCR擴(kuò)增出目的片段掌实，T7E1限制性酶切法檢測基因打靶情況，隨后進(jìn)行測序分析確定編輯效果邦马。通過O-dianisidine染色分析血紅蛋白合成情況贱鼻，中性粒細(xì)胞染色分析中性粒細(xì)胞生成情況，利用Real-time PCR檢測p53埃仪、mdm2即寡、hsp70和造血相關(guān)轉(zhuǎn)錄因子基因的表達(dá)變化验庙。結(jié)果 成功制備了rpl15 gRN......
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