An optimized FACS-free single-nucleus RNA sequencing (snRNA-seq) method for plant science research

生物 拟南芥 核糖核酸 计算生物学 基因 小核RNA 基因表达 单元格排序 遗传学 细胞生物学 细胞 非编码RNA 突变体
作者
Peng Wang,Caiyao Zhao,Sunhuan Xiang,Kunyu Duan,Xiaoli Chen,Xing Guo,Sunil Kumar Sahu
出处
期刊:Plant Science [Elsevier]
卷期号:326: 111535-111535 被引量:12
标识
DOI:10.1016/j.plantsci.2022.111535
摘要

Recently, single-cell RNA sequencing (scRNA-seq) provides unprecedented power for accurately understanding gene expression regulatory mechanisms. However, scRNA-seq studies have limitations in plants, due to difficulty in protoplast isolation that requires enzymatic digestion of the cell walls from various plant tissues. Therefore, to overcome this problem, we developed a nuclei isolation approach that does not rely on Fluorescence Activated Cell Sorting (FACS). We validated the robustness of the FACS-free single-nucleus RNA sequencing (snRNA-seq) methodology in mature Arabidopsis plant tissue by comparing it to scRNA-seq results based on protoplasts extracted from the same batch of leaf materials. Sequencing results demonstrated the high quality of snRNA-seq data, as well as its utility in cell type classification and marker gene identification. This approach also showed several advantages, including the ability to use frozen samples, taking less suspension preparation time, and reducing biased cellular coverage and dissociation-induced transcriptional artifacts. Surprisingly, snRNA-seq detected two epidermal pavement cell clusters, while scRNA-seq only had one. Furthermore, we hypothesized that these two epidermal cells represent the top and lower epidermis based on differences in expression patterns of cluster-specific expressed genes. In summary, this study has advanced the application of snRNA-seq in Arabidopsis leaves and confirmed the advantages of snRNA-seq in plant research.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
3秒前
内向的小凡完成签到,获得积分10
3秒前
zhentg完成签到,获得积分0
3秒前
紫熊发布了新的文献求助20
3秒前
5秒前
elgar612发布了新的文献求助10
5秒前
whf完成签到,获得积分10
8秒前
10秒前
我是老大应助科研通管家采纳,获得10
10秒前
慕青应助科研通管家采纳,获得10
10秒前
NexusExplorer应助科研通管家采纳,获得10
10秒前
CodeCraft应助科研通管家采纳,获得10
10秒前
10秒前
爆米花应助科研通管家采纳,获得10
11秒前
11秒前
安紊完成签到,获得积分10
11秒前
14秒前
马慕蕊完成签到 ,获得积分20
14秒前
所所应助1762120采纳,获得10
17秒前
李健的小迷弟应助lubeiyu采纳,获得10
18秒前
NexusExplorer应助ice采纳,获得10
21秒前
21秒前
甦龘关注了科研通微信公众号
25秒前
小蘑菇应助飞天奶酪采纳,获得10
26秒前
27秒前
步步高完成签到,获得积分10
28秒前
情怀应助1762120采纳,获得10
30秒前
可爱的函函应助sujiaoziemo采纳,获得10
30秒前
大模型应助坚强的严青采纳,获得10
31秒前
红红酱发布了新的文献求助20
32秒前
circle完成签到,获得积分10
33秒前
东如海应助陈如采纳,获得10
38秒前
39秒前
sujiaoziemo给sujiaoziemo的求助进行了留言
41秒前
ikun完成签到,获得积分10
41秒前
45秒前
46秒前
47秒前
zz完成签到 ,获得积分10
49秒前
二猫完成签到,获得积分10
50秒前
高分求助中
Sustainability in Tides Chemistry 1500
TM 5-855-1(Fundamentals of protective design for conventional weapons) 1000
Threaded Harmony: A Sustainable Approach to Fashion 799
Livre et militantisme : La Cité éditeur 1958-1967 500
Retention of title in secured transactions law from a creditor's perspective: A comparative analysis of selected (non-)functional approaches 500
"Sixth plenary session of the Eighth Central Committee of the Communist Party of China" 400
Introduction to Modern Controls, with illustrations in MATLAB and Python 310
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3057395
求助须知:如何正确求助?哪些是违规求助? 2713838
关于积分的说明 7437746
捐赠科研通 2358991
什么是DOI,文献DOI怎么找? 1249646
科研通“疑难数据库(出版商)”最低求助积分说明 607222
版权声明 596328