Quantifying tagged mRNA export flux via nuclear pore complexes in single live cells

被引：0
作者：
Jing, Yueyue [1 ]
Lv, Yilin [2 ,3 ]
Ye, Jingya [4 ]
Yao, Longfang [1 ]
Chen, Liwen [1 ]
Mi, Lan [1 ]
Fei, Yiyan [1 ]
Yu, Yao [2 ,3 ]
Dong, Biao [4 ]
Lv, Hong [2 ,3 ]
Ma, Jiong [1 ,5 ,6 ]
机构：
[1] Fudan Univ, Shanghai Engn Res Ctr Ultra Precis Opt Mfg, Key Lab Micro & Nano Photon Struct,Minist Educ, Dept Opt Sci & Engn,Green Photoelect Platform, Shanghai, Peoples R China
[2] Fudan Univ, Sch Life Sci, State Key Lab Genet Engn, Shanghai 200438, Peoples R China
[3] Shanghai Engn Res Ctr Ind Microorganisms, Shanghai 200438, Peoples R China
[4] Sichuan Univ, West China Hosp, Natl Clin Res Ctr Geriatr, Chengdu 610041, Sichuan, Peoples R China
[5] Fudan Univ, Acad Engineer & Technol, Inst Biomed Engn & Technol, Shanghai, Peoples R China
[6] Fudan Univ, Sch Life Sci, Multiscale Res Inst Complex Syst MRICS, Shanghai Engn Res Ctr Ind Microorganisms, Shanghai, Peoples R China
来源：
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2021年 / 545卷
基金：
中国国家自然科学基金; 国家重点研发计划;
关键词：
Gene expression monitoring; RNA Transport; Nuclear pore; Fluorescence microscopy; Modeling;
D O I：
10.1016/j.bbrc.2021.01.049
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The mRNA export flux through nuclear pore complexes (NPC) changes under DNA manipulation and hence affects protein translation. However, monitoring the flux of a specific mRNA in single live cell is beyond reach of traditional techniques. We developed a fluorescence-based detection method for measuring the export flux of mRNA through NPC in single live cell using a snapshot image, which had been tested on exogenous genes' expression in HeLa cells, with transfection or infection, and endogenous genes' expression in yeast cells, during incubation and carbon catabolite repression. With its speediness, explicitness and noninvasiveness, we believe that it would be valuable in direct monitoring of gene behavior, and the understanding of gene regulation at a single cell level. (c) 2021 Elsevier Inc. All rights reserved.
引用
收藏
页码：138 / 144
页数：7
相关论文
共 30 条
  • [1] Elimination of Toxic Microsatellite Repeat Expansion RNA by RNA-Targeting Cas9
    Batra, Ranjan
    Nelles, David A.
    Pirie, Elaine
    Blue, Steven M.
    Marina, Ryan J.
    Wang, Harrison
    Chaim, Isaac A.
    Thomas, James D.
    Zhang, Nigel
    Nguyen, Vu
    Aigner, Stefan
    Markmiller, Sebastian
    Xia, Guangbin
    Corbett, Kevin D.
    Swanson, Maurice S.
    Yeo, Gene W.
    [J]. CELL, 2017, 170 (05) : 899 - +
  • [2] Localization of ASH1 mRNA particles in living yeast
    Bertrand, E
    Chartrand, P
    Schaefer, M
    Shenoy, SM
    Singer, RH
    Long, RM
    [J]. MOLECULAR CELL, 1998, 2 (04) : 437 - 445
  • [3] Variability of the Reverse Transcription Step: Practical Implications
    Bustin, Stephen
    Dhillon, Harvinder S.
    Kirvell, Sara
    Greenwood, Christina
    Parker, Michael
    Shipley, Gregory L.
    Nolan, Tania
    [J]. CLINICAL CHEMISTRY, 2015, 61 (01) : 202 - 212
  • [4] Processing of recombinant AAV genomes occurs in specific nuclear structures that overlap with foci of DNA-damage-response proteins
    Cervelli, Tiziana
    Palacios, Jose Alejandro
    Zentilin, Lorena
    Mano, Miguel
    Schwartz, Rachel A.
    Weitzman, Matthew D.
    Giacca, Mauro
    [J]. JOURNAL OF CELL SCIENCE, 2008, 121 (03) : 349 - 357
  • [5] Enhanced Factor VIII Heavy Chain for Gene Therapy of Hemophilia A
    Chen, Lingxia
    Lu, Hui
    Wang, Jinhui
    Sarkar, Rita
    Yang, Xiao
    Wang, Hongli
    High, Katherine A.
    Xiao, Weidong
    [J]. MOLECULAR THERAPY, 2009, 17 (03) : 417 - 424
  • [6] Visualizing RNA dynamics in live cells with bright and stable fluorescent RNAs
    Chen, Xianjun
    Zhang, Dasheng
    Su, Ni
    Bao, Bingkun
    Xie, Xin
    Zuo, Fangting
    Yang, Lipeng
    Wang, Hui
    Jiang, Li
    Lin, Qiuning
    Fang, Mengyue
    Li, Ningfeng
    Hua, Xin
    Chen, Zhengda
    Bao, Chunyan
    Xu, Jinjin
    Du, Wenli
    Zhang, Lixin
    Zhao, Yuzheng
    Zhu, Linyong
    Loscalzo, Joseph
    Yang, Yi
    [J]. NATURE BIOTECHNOLOGY, 2019, 37 (11) : 1287 - +
  • [7] Gichner T, 1982, BIOL PLANTARUM, V24, P340
  • [8] In-cell identification and measurement of RNA-protein interactions
    Graindorge, Antoine
    Pinheiro, Ines
    Nawrocka, Anna
    Mallory, Allison C.
    Tsvetkov, Peter
    Gil, Noa
    Carolis, Carlo
    Buchholz, Frank
    Ulitsky, Igor
    Heard, Edith
    Taipale, Mikko
    Shkumatava, Alena
    [J]. NATURE COMMUNICATIONS, 2019, 10 (1)
  • [9] In vivo imaging of labelled endogenous β-actin mRNA during nucleocytoplasmic transport
    Grunwald, David
    Singer, Robert H.
    [J]. NATURE, 2010, 467 (7315) : 604 - U128
  • [10] DIFFERENTIALLY REGULATED MALATE SYNTHASE GENES PARTICIPATE IN CARBON AND NITROGEN-METABOLISM OF SACCHAROMYCES-CEREVISIAE
    HARTIG, A
    SIMON, MM
    SCHUSTER, T
    DAUGHERTY, JR
    YOO, HS
    COOPER, TG
    [J]. NUCLEIC ACIDS RESEARCH, 1992, 20 (21) : 5677 - 5686
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