硅壳结构由基因决定，明晰基因与硅壳特性之关联，是实现硅壳精准制造的前提。体外研究表明，聚阳离子多肽（Silaffins）可在不同尺度上影响硅聚合形态，表明其在硅壳合成中发挥重要作用。Silaffins氨基酸序列缺乏保守性，且经历复杂的翻译后修饰；因此，解析Silaffins基因极其困难，体内功能验证难以开展。单细胞拉曼技术，可能为Silaffins编码基因解析提供机会；而过量表达、RNA干扰、Crispr/Cas9等基因编辑工具，能够为基因功能本体验证创造条件。因此，本项目拟在小环藻中建立单细胞拉曼光谱“表型-分选-测序”技术，以获取Silaffins编码基因信息，进而通过不同基因编辑手段，探讨如下科学问题：Silaffins各成员在胞内是否在不同空间尺度上发挥功能？Silaffins是否是硅壳种间特异结构的决定因素？以Silaffins为切入点进行基因编辑，能够对硅壳结构产生何种改变？

Genes determine the architectural structure of diatom frustule. Therefore, clarification of the relationship between the gene and the frustule structure is the premise to realize the precise modification of frustule by genetic engineering. In vitro studies have widely shown that the Silaffins can affect the morphology of silica condensation on different scales, demonstrating that they must play an important roles in the biosynthesis of frustule. However, the amino acid sequences of Silaffins lack conservatism, and all of them undergo complex post-translational modification processes, so it is extremely difficult to identify corresponding genes of Silaffins. The development of single cell Raman spectroscopy provides an opportunity for the determination of genes encoding Silaffins, and the establishment genetic editing systems (e.g., overexpression, RNA interference, Crispr/Cas9) can conduct functional analysis of genes efficiently. Above all, this project intends to establish a technological platform of “Phenotypic examination-sorting-sequencing” based on single cell resonance Raman spectra to obtain the genes encoding Silaffins, and then, to solve the key scientific problems in Cyclotella cryptica as follows, through multiple gene editing means: Whether various members of Silaffins contribute to the frustule formation on different scales? Do the species-specific Silaffins determine the species-specific structure of frustules? Whether it is possible to modify the frustule structure in various scales through gene editing?