Zhao Xinlu, Sun Shiming, Yu Wenhao, Zhu Wenqi, Zhao Zihan, Zhou Yiqi, Ding Xiuheng, Fang Nan, Yang Rong, Li Jie P
State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center (ChemBIC), School of Chemistry and Chemical Engineering, Nanjing University Nanjing Jiangsu China
Singleron Biotechnologies Nanjing Jiangsu China.
RSC Chem Biol. 2022 May 31;3(8):1052-1060. doi: 10.1039/d2cb00046f. eCollection 2022 Aug 3.
Click chemistry-enabled DNA barcoding of cells provides a universal strategy for sample multiplexing in single-cell RNA-seq (scRNA-seq). However, current ClickTags are limited to fixed samples as they only label cells efficiently in methanol. Herein, we report the development of a new protocol for barcoding live cells with improved ClickTags. The optimized reactions barcoded live cells without perturbing their physiological states, which allowed sample multiplexing of live cells in scRNA-seq. The general applicability of this protocol is demonstrated in diversified types of samples, including murine and human primary samples. Up to 16 samples across these two species are successfully multiplexed and demultiplexed with high consistency. The wide applications of this method could help to increase throughput, reduce cost and remove the batch effect in scRNA-seq, which is especially valuable for studying clinical samples from a large cohort.
基于点击化学的细胞DNA条形码技术为单细胞RNA测序（scRNA-seq）中的样本多重分析提供了一种通用策略。然而，目前的点击标签仅限于固定样本，因为它们仅在甲醇中能有效地标记细胞。在此，我们报告了一种使用改进的点击标签对活细胞进行条形码标记的新方案。优化后的反应能够对活细胞进行条形码标记，同时不干扰其生理状态，从而实现了scRNA-seq中活细胞的样本多重分析。该方案在多种类型的样本中都得到了验证，包括小鼠和人类的原代样本。这两个物种的多达16个样本成功地进行了多重分析和解复用，且具有高度的一致性。该方法的广泛应用有助于提高通量、降低成本并消除scRNA-seq中的批次效应，这对于研究来自大量队列的临床样本尤为重要。
