Fast and Efficient CRISPR/Cas9 Genome Editing In Vivo Enabled by Bioreducible Lipid and Messenger RNA Nanoparticles
Fast and Efficient CRISPR/Cas9 Genome Editing In Vivo Enabled by Bioreducible Lipid and Messenger RNA Nanoparticles
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通过生物可还原脂质和信使 RNA 纳米颗粒实现快速高效的体内 CRISPR/Cas9 基因组编辑
DOI：
10.1002/adma.201902575
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发表时间：
2019-08-01
影响因子：
29.4
通讯作者：
Wang, Ming
中科院分区：
文献类型：
--
作者：
Liu, Ji;Chang, Jin;Wang, Ming
拓宽CRISPR/Cas9（成簇规则间隔短回文重复序列（CRISPR）相关蛋白9）基因组编辑技术的生物医学应用的主要挑战是将Cas9核酸酶和单向导RNA（sgRNA）递送到特定细胞和器官中。报道了通过生物可还原脂质/Cas9信使RNA（mRNA）纳米颗粒实现的体外和体内有效且非常快速的CRISPR/Cas9基因组编辑。BAMEA-O 16 B是一种整合有二硫键的脂质纳米颗粒，可以有效地将Cas9 mRNA和sgRNA递送到细胞中，同时响应于还原性细胞内环境释放RNA，以在mRNA递送后24小时内进行基因组编辑。证明使用BAMEA-O 16 B同时递送Cas9 mRNA和sgRNA敲除人胚肾细胞的绿色荧光蛋白（GFP）表达，效率高达90%。此外，静脉内注射BAMEA-O 16 B/Cas9 mRNA/sgRNA纳米颗粒有效地在肝细胞中积累，并将小鼠血清中的前蛋白转化酶枯草杆菌蛋白酶/kexin 9型水平敲低至未处理的20%。领先的脂质纳米颗粒BAMEA-O 16 B代表了迄今为止报道的最有效的CRISPR/Cas9递送纳米载体之一，它可以进一步拓宽mRNA和CRISPR/Cas9技术的治疗前景。
A main challenge to broaden the biomedical application of CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat (CRISPR) associated protein 9) genome editing technique is the delivery of Cas9 nuclease and single-guide RNA (sgRNA) into the specific cell and organ. An effective and very fast CRISPR/Cas9 genome editing in vitro and in vivo enabled by bioreducible lipid/Cas9 messenger RNA (mRNA) nanoparticle is reported. BAMEA-O16B, a lipid nanoparticle integrated with disulfide bonds, can efficiently deliver Cas9 mRNA and sgRNA into cells while releasing RNA in response to the reductive intracellular environment for genome editing as fast as 24 h post mRNA delivery. It is demonstrated that the simultaneous delivery of Cas9 mRNA and sgRNA using BAMEA-O16B knocks out green fluorescent protein (GFP) expression of human embryonic kidney cells with efficiency up to 90%. Moreover, the intravenous injection of BAMEA-O16B/Cas9 mRNA/sgRNA nanoparticle effectively accumulates in hepatocytes, and knocks down proprotein convertase subtilisin/kexin type 9 level in mouse serum down to 20% of nontreatment. The leading lipid nanoparticle, BAMEA-O16B, represents one of the most efficient CRISPR/Cas9 delivery nanocarriers reported so far, and it can broaden the therapeutic promise of mRNA and CRISPR/Cas9 technique further.