近日，美国加州大学旧金山分校Zev J. Gartner、Eric D. Chow及其课题组提出了使用脂质标记进行单细胞RNA多路测序的技术。 该项研究成果发表在2019年7月出版的《Nature Methods》上。

研究团队开发了一种新的多路单细胞RNA测序技术，研究者利用脂质标记实现单细胞和单核RNA测序。该多路测序试剂通过对可接近质膜进行标记，实现对任意物种的任意类型细胞或细胞核进行条形码标记。该方法涉及最小化样本处理，从而维持细胞活力和内源性基因表达模式。研究者依据该多路测序条形码的丰度将细胞分组，通过进行双重确认和回收RNA含量较低的细胞，从而提高获取数据的质量，避免数据在标准质量控制工作流程中丢失。研究者对原发性人乳腺上皮细胞、人源肿瘤异体种移植小鼠模型（三阴性乳腺癌）分离出的多重冷冻保存肿瘤组织和转移部位，进行96-plex干扰实验，使用MULTI-seq跟踪T细胞活化的动态过程。

据了解，单细胞RNA测序中，样本多路使用能够有效降低检测成本和排除双细胞假象。然而，目前的样本条形码标记技术还不具备很好的普遍适用性与可扩展性。

附：英文原文

Title: MULTI-seq: sample multiplexing for single-cell RNA sequencing using lipid-tagged indices

Author: Christopher S. McGinnis, David M. Patterson, Juliane Winkler, Daniel N. Conrad, Marco Y. Hein, Vasudha Srivastava, Jennifer L. Hu, Lyndsay M. Murrow, Jonathan S. Weissman, Zena Werb, Eric D. Chow, Zev J. Gartner

Issue&Volume: Volume 16 Issue 7, July 2019

Abstract: Sample multiplexing facilitates scRNA-seq by reducing costs and identifying artifacts such as cell doublets. However, universal and scalable sample barcoding strategies have not been described. We therefore developed MULTI-seq: multiplexing using lipid-tagged indices for single-cell and single-nucleus RNA sequencing. MULTI-seq reagents can barcode any cell type or nucleus from any species with an accessible plasma membrane. The method involves minimal sample processing, thereby preserving cell viability and endogenous gene expression patterns. When cells are classified into sample groups using MULTI-seq barcode abundances, data quality is improved through doublet identification and recovery of cells with low RNA content that would otherwise be discarded by standard quality-control workflows. We use MULTI-seq to track the dynamics of T-cell activation, perform a 96-plex perturbation experiment with primary human mammary epithelial cells and multiplex cryopreserved tumors and metastatic sites isolated from a patient-derived xenograft mouse model of triple-negative breast cancer.

DOI: 10.1038/s41592-019-0433-8

Source: https://www.nature.com/articles/s41592-019-0433-8