Immunocompromised Cas9 transgenic mice for rapid in vivo assessment of host factors involved in highly pathogenic virus infection.
Immunocompromised Cas9 transgenic mice for rapid in vivo assessment of host factors involved in highly pathogenic virus infection.
复制标题
DOI：
10.1016/j.omtm.2021.09.012
复制
发表时间：
2021-12-10
期刊：
影响因子：
--
通讯作者：
Negrete OA
中科院分区：
文献类型：
--
作者：
Collette N;Dhungel P;Lund SJ;Schwedler JL;Saada EA;Light YK;Sinha A;Schoeniger JS;Negrete OA
靶向宿主因子的抗病毒开发提供了几个潜在的优势，传统的对策，包括广谱活性和耐药性的预防。宿主因子在动物模型中的表征提供了它们参与疾病发病机制的强有力证据，但是对基因列表进行高通量体内分析的可行性是有问题的。为了开始解决体内筛选候选宿主因子的挑战，我们将CRISPR-Cas9基因组编辑的进展与用于研究高致病性病毒的免疫受损小鼠模型相结合。携带组成型表达的Cas9等位基因（Cas9 tg/tg）的转基因小鼠（具有或不具有I型干扰素受体的敲除）用于使用Invivofectamine 3.0（一种简单且易于使用的脂质纳米颗粒试剂）优化CRISPR单向导RNA（sgRNA）的体内递送。Invivofectamine 3.0介导的肝脏特异性编辑在平均74%的肝细胞中去除关键埃博拉病毒宿主因子尼曼-匹克病C1型的活性，保护免疫受损的Cas9 tg/tg小鼠免受致命的替代埃博拉病毒感染。我们设想，免疫受损的Cas9 tg/tg小鼠与直接的sgRNA体内递送相结合，将能够在肝脏和其他器官中进行有效的宿主因子功能丧失筛选，以快速研究它们对病毒发病机制的影响，并帮助启动针对新出现的病原体的广谱宿主导向疗法的开发。Collette等人报告使用Invivofectamine 3.0（一种易于使用的脂质纳米颗粒试剂）在Cas9转基因小鼠中优化CRISPR sgRNA的体内递送。他们强调了免疫受损Cas9转基因小鼠中Npc 1的优化肝脏特异性编辑产生的部分敲除表型，可以完全保护免受致命的替代埃博拉病毒的攻击。
Targeting host factors for anti-viral development offers several potential advantages over traditional countermeasures that include broad-spectrum activity and prevention of resistance. Characterization of host factors in animal models provides strong evidence of their involvement in disease pathogenesis, but the feasibility of performing high-throughput in vivo analyses on lists of genes is problematic. To begin addressing the challenges of screening candidate host factors in vivo, we combined advances in CRISPR-Cas9 genome editing with an immunocompromised mouse model used to study highly pathogenic viruses. Transgenic mice harboring a constitutively expressed Cas9 allele (Cas9tg/tg) with or without knockout of type I interferon receptors served to optimize in vivo delivery of CRISPR single-guide RNA (sgRNA) using Invivofectamine 3.0, a simple and easy-to-use lipid nanoparticle reagent. Invivofectamine 3.0-mediated liver-specific editing to remove activity of the critical Ebola virus host factor Niemann-Pick disease type C1 in an average of 74% of liver cells protected immunocompromised Cas9tg/tg mice from lethal surrogate Ebola virus infection. We envision that immunocompromised Cas9tg/tg mice combined with straightforward sgRNA in vivo delivery will enable efficient host factor loss-of-function screening in the liver and other organs to rapidly study their effects on viral pathogenesis and help initiate development of broad-spectrum, host-directed therapies against emerging pathogens. Collette et al. report optimized in vivo delivery of CRISPR sgRNA in Cas9 transgenic mice using Invivofectamine 3.0, an easy-to-use lipid nanoparticle reagent. They highlight a partial knockout phenotype resulting from optimized liver-specific editing of Npc1 in immunocompromised Cas9 transgenic mice as being completely protective against lethal surrogate Ebola virus challenge.
登录
查看更多内容
影响因子：
6.4
作者：
Jones, Steven M.;Stroeher, Ute;Feldmann, Heinz
通讯作者：
Feldmann, Heinz
影响因子：
3.7
作者：
Smith, Darci R.;Steele, Keith E.;Hensley, Lisa E.
通讯作者：
Hensley, Lisa E.
影响因子：
7.7
作者：
Ng, Melinda;Ndungo, Esther;Chandran, Kartik
通讯作者：
Chandran, Kartik
DOI：
10.1038/nrd.2017.162
发表时间：
2018-01
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
Kaufmann SHE;Dorhoi A;Hotchkiss RS;Bartenschlager R
通讯作者：
Bartenschlager R
影响因子：
6.4
作者：
Marzi, Andrea;Kercher, Lisa;Feldmann, Heinz
通讯作者：
Feldmann, Heinz