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Measuring DNA Hybridization Kinetics in Live Cells Using a Time-Resolved 3D Single-Molecule Tracking Method.
Chen, Yuan-I; Chang, Yin-Jui; Nguyen, Trung Duc; Liu, Cong; Phillion, Stephanie; Kuo, Yu-An; Vu, Huong T; Liu, Angela; Liu, Yen-Liang; Hong, Soonwoo; Ren, Pengyu; Yankeelov, Thomas E; Yeh, Hsin-Chih.
Afiliação
  • Chen YI; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Chang YJ; Department of Mechanical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Nguyen TD; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Liu C; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Phillion S; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Kuo YA; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Vu HT; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Liu A; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Liu YL; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Hong S; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Ren P; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Yankeelov TE; Department of Biomedical Engineering , University of Texas at Austin , Austin , Texas 78712 , United States.
  • Yeh HC; Oden Institute for Computational Engineering and Sciences, University of Texas at Austin , Austin , Texas 78712 , United States.
J Am Chem Soc ; 141(40): 15747-15750, 2019 10 09.
Article em En | MEDLINE | ID: mdl-31509386
ABSTRACT
Single-molecule detection enables direct characterization of annealing/melting kinetics of nucleic acids without the need for synchronization of molecular states, but the current experiments are not carried out in a native cellular context. Here we describe an integrated 3D single-molecule tracking and lifetime measurement method that can follow individual DNA molecules diffusing inside a mammalian cell and observe multiple annealing and melting events on the same molecules. By comparing the hybridization kinetics of the same DNA strand in vitro, we found the association constants can be 13- to 163-fold higher in the molecular crowding cellular environment.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA / Imagem Individual de Molécula / Conformação de Ácido Nucleico / Hibridização de Ácido Nucleico Tipo de estudo: Health_economic_evaluation Idioma: En Ano de publicação: 2019 Tipo de documento: Article
Texto completo
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XML
PubMed Links
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA / Imagem Individual de Molécula / Conformação de Ácido Nucleico / Hibridização de Ácido Nucleico Tipo de estudo: Health_economic_evaluation Idioma: En Ano de publicação: 2019 Tipo de documento: Article