Department of Cellular and Developmental Biology of Plants, University of Bielefeld, Universitätsstr. 25, 33615 Bielefeld, Germany.
Genes (Basel). 2023 Jul 1;14(7):1389. doi: 10.3390/genes14071389.
The multicellular green alga has emerged as a valuable model organism for investigating various aspects of multicellularity and cellular differentiation, photoreception and phototaxis, cell division, biogenesis of the extracellular matrix and morphogenetic movements. While a range of molecular tools and bioinformatics resources have been made available for exploring these topics, the establishment of cell type-specific promoters in has not been achieved so far. Therefore, here, we conducted a thorough screening of transcriptome data from RNA sequencing analyses of in order to identify potential cell type-specific promoters. Eventually, we chose two putative strong and cell type-specific promoters, with one exhibiting specific expression in reproductive cells (gonidia), the promoter, and the other in somatic cells, the promoter. After cloning both promoter regions, they were introduced upstream of a luciferase reporter gene. By using particle bombardment, the DNA constructs were stably integrated into the genome of . The results of the expression analyses, which were conducted at both the transcript and protein levels, demonstrated that the two promoters drive cell type-specific expression in their respective target cell types. Transformants with considerably diverse expression levels of the chimeric genes were identifiable. In conclusion, the screening and analysis of transcriptome data from RNA sequencing allowed for the identification of potential cell type-specific promoters in . Reporter gene constructs demonstrated the actual usability of two promoters. The investigated and promoters were proven to be potent molecular tools for genetic engineering in .
多细胞绿藻已成为研究多细胞性和细胞分化、光感受和趋光性、细胞分裂、细胞外基质生物发生和形态发生运动等各个方面的有价值的模式生物。虽然已经有一系列的分子工具和生物信息学资源可用于探索这些主题，但到目前为止，尚未在 中建立细胞类型特异性启动子。因此，在这里，我们对 的 RNA 测序分析的转录组数据进行了全面筛选，以鉴定潜在的细胞类型特异性启动子。最终，我们选择了两个假定的强且细胞类型特异性启动子，一个在生殖细胞（生殖细胞）中特异性表达，即 启动子，另一个在体细胞中表达，即 启动子。克隆两个启动子区域后，它们被引入到荧光素酶报告基因的上游。通过使用粒子轰击，将 DNA 构建体稳定整合到 的基因组中。在转录和蛋白质水平上进行的表达分析结果表明，这两个启动子在各自的靶细胞类型中驱动细胞类型特异性表达。可以识别出具有不同嵌合基因表达水平的转化体。总之，从 RNA 测序的转录组数据筛选和分析可鉴定 的潜在细胞类型特异性启动子。报告基因构建体证明了两个启动子在 中的实际可用性。所研究的 和 启动子被证明是 在 中进行遗传工程的有效分子工具。
