Systematic engineering of virus-like particles to identify self-assembly rules for shifting particle size
Systematic engineering of virus-like particles to identify self-assembly rules for shifting particle size
复制标题
病毒样颗粒的系统工程，以确定改变颗粒尺寸的自组装规则
DOI：
10.1016/j.virol.2023.01.002
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发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
Tullman-Ercek, Danielle
中科院分区：
文献类型：
--
作者：
Ikwuagwu, Bon;Hartman, Emily;Mills, Carolyn E.;Tullman-Ercek, Danielle
病毒样颗粒（vlp）是一种很有前途的生物材料支架以及诊断和治疗应用。然而，有一些关键的挑战需要解决，比如为不同的用例设计可替换大小的能力。为此，我们在外壳蛋白的两个关键残基上创建了MS2 VLP变体库，这些残基对控制VLP的大小和几何形状很重要。通过采用系统诱变方法，结合基于大小的选择和高通量测序作为读出，我们对MS2外壳蛋白中控制MS2 VLPs大小变化的两个残基进行了定量评估。然后，我们将该策略应用于Qβ VLP （MS2同源物）中的等效残基，并证明了类似的残基对也能够影响Qβ VLP的大小和形状。这些结果强调了适应度景观在识别装配的关键特征方面的力量。
Virus-like particles (VLPs) are promising scaffolds for biomaterials as well as diagnostic and therapeutic applications. However, there are some key challenges to be solved, such as the ability to engineer alternate sizes for varied use cases. To this end, we created a library of MS2 VLP variants at two key residues in the coat protein which have been implicated as important to controlling VLP size and geometry. By adapting a method for systematic mutagenesis coupled with size-based selections and high-throughput sequencing as a readout, we developed a quantitative assessment of two residues in MS2 coat protein that govern the size shift in MS2 VLPs. We then applied the strategy to the equivalent residues in Qβ VLPs, an MS2 homolog, and demonstrate that the analogous pair of residues are also able to impact Qβ VLP size and shape. These results underscore the power of fitness landscapes in identifying critical features for assembly.