Rapid assessment of SARS-CoV-2-evolved variants using virus-like particles.
Rapid assessment of SARS-CoV-2-evolved variants using virus-like particles.
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DOI：
10.1126/science.abl6184
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发表时间：
2021-12-24
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为了开发针对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）和新出现的变体的治疗方法，了解病毒生物学和突变的影响非常重要。然而，这是具有挑战性的，因为活病毒只能在少数符合严格安全标准的实验室中进行研究。Syed等人描述了一种病毒样颗粒（VLP），它包含四种SARS-CoV-2结构蛋白，但不是包装病毒RNA，而是包装表达报告蛋白的信使RNA（mRNA）（参见约翰逊和Menachery的《透视》）。受体细胞中表达的报告基因的量取决于生产细胞中包装和组装的效率以及进入受体细胞的效率。在更具传播性的变体中发现的核衣壳蛋白的突变增加mRNA包装和表达。VLP为研究结构蛋白突变的影响和筛选治疗方法提供了平台。-VV SARS-CoV-2样颗粒能够快速安全地分析病毒变体。确定为什么新的严重急性呼吸综合征冠状病毒2（SARS-CoV-2）变体表现出更好的适应性的努力仅限于使用S-假型颗粒分析刺突（S）蛋白的突变。在这项研究中，我们表明SARS-CoV-2病毒样颗粒（SC2-VLP）可以包装和传递外源性转录本，从而能够分析所有结构蛋白和病毒生命周期中多个步骤的突变。在SC2-VLP中，普遍存在于更可传播的变体中的四个核衣壳（N）突变独立地增加信使RNA递送和表达约10倍，并且在反向遗传学模型中，丝氨酸-202 →精氨酸（S202 R）和精氨酸-203 →甲硫氨酸（R203 M）突变各自产生>50倍的病毒。SC2-VLP提供了一个平台，用于快速检测生物安全3级环境之外的病毒变体，并证明N突变和颗粒组装是可以解释变体传播增加的机制，包括B.1.617.2（Delta，包含R203 M突变）。
To develop therapies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and emerging variants, it is important to understand the viral biology and the effect of mutations. However, this is challenging because live virus can only be studied in a few laboratories that meet stringent safety standards. Syed et al. describe a virus-like particle (VLP) that comprises the four SARS-CoV-2 structural proteins, but instead of packaging viral RNA, it packages messenger RNA (mRNA) that expresses a reporter protein (see the Perspective by Johnson and Menachery). The amount of reporter expressed in receiver cells depends on the efficiency of packaging and assembly in the producer cells and the efficiency of entry into receiver cells. Mutations in the nucleocapsid protein that are found in more transmissible variants increase mRNA packaging and expression. The VLPs provide a platform for studying the effect of mutations in the structural proteins and for screening therapeutics. —VV SARS-CoV-2–like particles enable rapid and safe analysis of viral variants. Efforts to determine why new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants demonstrate improved fitness have been limited to analyzing mutations in the spike (S) protein with the use of S-pseudotyped particles. In this study, we show that SARS-CoV-2 virus-like particles (SC2-VLPs) can package and deliver exogenous transcripts, enabling analysis of mutations within all structural proteins and at multiple steps in the viral life cycle. In SC2-VLPs, four nucleocapsid (N) mutations found universally in more-transmissible variants independently increased messenger RNA delivery and expression ~10-fold, and in a reverse genetics model, the serine-202→arginine (S202R) and arginine-203→methionine (R203M) mutations each produced >50 times as much virus. SC2-VLPs provide a platform for rapid testing of viral variants outside of a biosafety level 3 setting and demonstrate N mutations and particle assembly to be mechanisms that could explain the increased spread of variants, including B.1.617.2 (Delta, which contains the R203M mutation).
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影响因子：
9.8
作者：
Rihn SJ;Merits A;Bakshi S;Turnbull ML;Wickenhagen A;Alexander AJT;Baillie C;Brennan B;Brown F;Brunker K;Bryden SR;Burness KA;Carmichael S;Cole SJ;Cowton VM;Davies P;Davis C;De Lorenzo G;Donald CL;Dorward M;Dunlop JI;Elliott M;Fares M;da Silva Filipe A;Freitas JR;Furnon W;Gestuveo RJ;Geyer A;Giesel D;Goldfarb DM;Goodman N;Gunson R;Hastie CJ;Herder V;Hughes J;Johnson C;Johnson N;Kohl A;Kerr K;Leech H;Lello LS;Li K;Lieber G;Liu X;Lingala R;Loney C;Mair D;McElwee MJ;McFarlane S;Nichols J;Nomikou K;Orr A;Orton RJ;Palmarini M;Parr YA;Pinto RM;Raggett S;Reid E;Robertson DL;Royle J;Cameron-Ruiz N;Shepherd JG;Smollett K;Stewart DG;Stewart M;Sugrue E;Szemiel AM;Taggart A;Thomson EC;Tong L;Torrie LS;Toth R;Varjak M;Wang S;Wilkinson SG;Wyatt PG;Zusinaite E;Alessi DR;Patel AH;Zaid A;Wilson SJ;Mahalingam S
通讯作者：
Mahalingam S
影响因子：
64.8
作者：
Plante JA;Liu Y;Liu J;Xia H;Johnson BA;Lokugamage KG;Zhang X;Muruato AE;Zou J;Fontes-Garfias CR;Mirchandani D;Scharton D;Bilello JP;Ku Z;An Z;Kalveram B;Freiberg AN;Menachery VD;Xie X;Plante KS;Weaver SC;Shi PY
通讯作者：
Shi PY
影响因子：
16.6
作者：
Li B;Deng A;Li K;Hu Y;Li Z;Shi Y;Xiong Q;Liu Z;Guo Q;Zou L;Zhang H;Zhang M;Ouyang F;Su J;Su W;Xu J;Lin H;Sun J;Peng J;Jiang H;Zhou P;Hu T;Luo M;Zhang Y;Zheng H;Xiao J;Liu T;Tan M;Che R;Zeng H;Zheng Z;Huang Y;Yu J;Yi L;Wu J;Chen J;Zhong H;Deng X;Kang M;Pybus OG;Hall M;Lythgoe KA;Li Y;Yuan J;He J;Lu J
通讯作者：
Lu J
DOI：
10.15252/embj.2020106478
发表时间：
2020-12-15
期刊：
The EMBO journal
影响因子：
--
作者：
Perdikari TM;Murthy AC;Ryan VH;Watters S;Naik MT;Fawzi NL
通讯作者：
Fawzi NL
DOI：
10.3390/pathogens9110912
发表时间：
2020-11-04
期刊：
Pathogens (Basel, Switzerland)
影响因子：
--
作者：
Kanakan A;Mishra N;Srinivasa Vasudevan J;Sahni S;Khan A;Sharma S;Pandey R
通讯作者：
Pandey R