Adaptive RNA editing in Cephalopods

我们在神经乌贼器官中应用了不同的单细胞RNA测序技术。其中一种技术使用10X基因组学和ONT或PacBio的长读测序相结合，从中我们观察到与细胞类型对应的3'端同源型特异性和rna编辑。但是，每个细胞的基因覆盖率和基因含量仍然很低。我们还观察到，头足类动物高度重复的基因组限制了整个基因同种型的测序，并产生了更短的序列和来自链入侵模式的显著水平的分子。我们已经开始使用基于平板的分选对单细胞进行测序，以解决我们目前的局限性。利用我们的红毛菊基因组，我修改了TSO-UMI引物，以减少链入侵的分子数量。结果，我们观察到的cDNA峰值比以前的技术要大得多，这表明在RT-PCR过程中成功地收集了新鲜细胞，并跳过了链入侵的分子。

We have applied different single-cell RNA sequencing technologies in neural squid organs. One of these technologies uses a combination of 10X genomics and long-read sequencing with ONT or PacBio, from which we observed isoform-specific and RNA-editing at the 3' end corresponding to a cell type. However, the gene coverage and gene content per cell were still very low. We also observed that the highly repetitive genome of cephalopods limits the sequencing of entire gene isoforms and generates shorter sequences and significant levels of molecules coming from strand-invasion patterns. We have started sequencing single cells using plate-based sorting to solve our current limitations. Using our Euprymna berryi genome, I modified TSO-UMI primers to reduce the number of molecules with strand invasion. As a result, we have observed much larger cDNA peaks than in the previous technologies, indicating a successful fresh cell collection and skipping molecules with strand invasion during the RT-PCR.