title: Single-cell mRNA transfection studies: delivery, kinetics and statistics by numbers.
abstract: In artificial gene delivery, messenger RNA (mRNA) is an attractive alternative to plasmid DNA (pDNA) since it does not require transfer into the cell nucleus. Here we show that, unlike for pDNA transfection, the delivery statistics and dynamics of mRNA-mediated expression are generic and predictable in terms of mathematical modeling. We measured the single-cell expression time-courses and levels of enhanced green fluorescent protein (eGFP) using time-lapse microscopy and flow cytometry (FC). The single-cell analysis provides direct access to the distribution of onset times, life times and expression rates of mRNA and eGFP. We introduce a two-step stochastic delivery model that reproduces the number distribution of successfully delivered and translated mRNA molecules and thereby the dose-response relation. Our results establish a statistical framework for mRNA transfection and as such should advance the development of RNA carriers and small interfering/micro RNA-based drugs.
  
  
  BACKGROUND
  This team of authors established a statistical framework for mRNA transfection by using a two-step stochastic delivery model that reproduces the number distribution of successfully delivered and translated mRNA molecules and thereby their dose-response relation. This study establishes a nice connection between theory and experimental planning and will aid the cellular delivery of mRNA molecules.
authors: Faculty of Physics and Center for NanoScience (CeNS), Ludwig-Maximilians-Universität, München, Germany.
authors: Carolin
authors: C
authors: Leonhardt
authors: Faculty of Physics and Center for NanoScience (CeNS), Ludwig-Maximilians-Universität, München, Germany.
authors: Gerlinde
authors: G
authors: Schwake
authors: Faculty of Physics and Center for NanoScience (CeNS), Ludwig-Maximilians-Universität, München, Germany.
authors: Tobias R
authors: TR
authors: Stögbauer
authors: Faculty of Physics and Center for NanoScience (CeNS), Ludwig-Maximilians-Universität, München, Germany.
authors: Susanne
authors: S
authors: Rappl
authors: Institut für theoretische Physik, Technische Universität Berlin, Berlin-Charlottenburg, Germany.
authors: Jan-Timm
authors: JT
authors: Kuhr
authors: Faculty of Physics and Center for NanoScience (CeNS), Ludwig-Maximilians-Universität, München, Germany.
authors: Thomas S
authors: TS
authors: Ligon
authors: Faculty of Physics and Center for NanoScience (CeNS), Ludwig-Maximilians-Universität, München, Germany. Electronic address: raedler@lmu.de.
authors: Joachim O
authors: JO
authors: Rädler
mesh_terms: D045744:Cell Line, Tumor; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007700:Kinetics; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics; D014162:Transfection / Q000379:methods*
chemical_list: D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Expression kinetics; Non-viral gene delivery; Pharmacokinetics; Single-cell studies; mRNA transfection