小柯机器人

德国亥姆霍兹感染研究中心（HZI）Chase L. Beisel团队近期取得重要工作进展，他们提出使用重编程tracrRNA在单个细菌细胞中记录RNA的策略。相关论文2023年1月5日在线发表于《自然—生物技术》杂志上。

据介绍，获取单个细胞的转录历史是一个挑战，细胞群落的功能异质性加剧了这一挑战。

研究人员利用重编程的tracrRNA（Rptrs）将选定的细胞转录物记录为单个活细菌细胞中存储的DNA编辑。Rptr被设计为与被识别转录物配对，将其转换为引导RNA。然后，引导RNA和Cas9碱基编辑器以引入的DNA靶点为目标。碱基编辑的程度可以在将来通过测序来读取。研究人员使用这种被称为TIGER（由基因编码记录推断的转录RNA）的方法来记录单个细菌细胞中的异源和内源性转录物。TIGER可以量化相对表达，区分单核苷酸差异，同时记录多个转录本并读出单细胞现象。研究人员进一步应用TIGER记录大肠杆菌中的代谢风险和抗生素耐药性以及沙门氏菌对宿主细胞的入侵。

总之，通过RNA记录，TIGER将当前的细胞状态与过去的转录状态联系起来，以破译单个细胞中复杂的细胞反应。

附：英文原文

Title: RNA recording in single bacterial cells using reprogrammed tracrRNAs

Author: Jiao, Chunlei, Reckstadt, Claas, Knig, Fabian, Homberger, Christina, Yu, Jiaqi, Vogel, Jrg, Westermann, Alexander J., Sharma, Cynthia M., Beisel, Chase L.

Issue&Volume: 2023-01-05

Abstract: Capturing an individual cell’s transcriptional history is a challenge exacerbated by the functional heterogeneity of cellular communities. Here, we leverage reprogrammed tracrRNAs (Rptrs) to record selected cellular transcripts as stored DNA edits in single living bacterial cells. Rptrs are designed to base pair with sensed transcripts, converting them into guide RNAs. The guide RNAs then direct a Cas9 base editor to target an introduced DNA target. The extent of base editing can then be read in the future by sequencing. We use this approach, called TIGER (transcribed RNAs inferred by genetically encoded records), to record heterologous and endogenous transcripts in individual bacterial cells. TIGER can quantify relative expression, distinguish single-nucleotide differences, record multiple transcripts simultaneously and read out single-cell phenomena. We further apply TIGER to record metabolic bet hedging and antibiotic resistance mobilization in Escherichia coli as well as host cell invasion by Salmonella. Through RNA recording, TIGER connects current cellular states with past transcriptional states to decipher complex cellular responses in single cells.

DOI: 10.1038/s41587-022-01604-8

Source: https://www.nature.com/articles/s41587-022-01604-8

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex