小柯机器人

美国斯坦福大学医学院Hanlee P. Ji.团队实现了单细胞水平癌突变及其转录表型的直接检测。2023年9月11日，国际学术期刊《自然-生物技术》发表了这一成果。

研究人员研发了一种名为TISCC-seq的高通量、多重单细胞技术；其使用CRISPR碱基编辑器在细胞中引入预先指定的突变，可直接在单个细胞中描绘它们的基因型并确定每个突变的转录表型。靶基因转录本长读长测序可识别工程化引入的突变，并且可通过短读长测序同时分析同一组细胞的转录组图谱。

通过整合数据，研究人员在单细胞分辨率水平确定了突变的基因型和表达表型。使用细胞系研究，研究人员设计和评估了100个TP53突变对基因表达的影响。基于单细胞基因表达，研究人员划分了具有显著功能的突变。

据悉，基因组测序研究揭示了存在于多种肿瘤中的许多突变类型，但确定由这些突变诱导的表型仍然是一个挑战。

附：英文原文

Title: Direct measurement of engineered cancer mutations and their transcriptional phenotypes in single cells

Author: Kim, Heon Seok, Grimes, Susan M., Chen, Tianqi, Sathe, Anuja, Lau, Billy T., Hwang, Gue-Ho, Bae, Sangsu, Ji, Hanlee P.

Issue&Volume: 2023-09-11

Abstract: Genome sequencing studies have identified numerous cancer mutations across a wide spectrum of tumor types, but determining the phenotypic consequence of these mutations remains a challenge. Here, we developed a high-throughput, multiplexed single-cell technology called TISCC-seq to engineer predesignated mutations in cells using CRISPR base editors, directly delineate their genotype among individual cells and determine each mutation’s transcriptional phenotype. Long-read sequencing of the target gene’s transcript identifies the engineered mutations, and the transcriptome profile from the same set of cells is simultaneously analyzed by short-read sequencing. Through integration, we determine the mutations’ genotype and expression phenotype at single-cell resolution. Using cell lines, we engineer and evaluate the impact of >100 TP53 mutations on gene expression. Based on the single-cell gene expression, we classify the mutations as having a functionally significant phenotype. Engineered cancer mutations are linked with phenotypes in a multiplexed single-cell technology.

DOI: 10.1038/s41587-023-01949-8

Source: https://www.nature.com/articles/s41587-023-01949-8

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex