title: Effect of Liposome Size on Internal RNA Replication Coupled with Replicase Translation.
abstract: Cell membranes inhibit the diffusion of intracellular materials, and compartment size can strongly affect the intracellular biochemical reactions. To assess the effect of the size of microcompartments on intracellular reactions, we constructed a primitive cell model consisting of giant liposomes and a translation-coupled RNA replication (TcRR) system. The RNA was replicated by Qβ replicase, which was translated from the RNA in giant liposomes encapsulating the cell-free translation system. A reporter RNA encoding the antisense strand of β-glucuronidase was introduced into the system to yield a TcRR read-out (green fluorescence). We demonstrate that TcRR was hardly detectable in larger liposomes (230 fL) but was more effective in smaller (7.7 fL) liposomes. Our experimental and theoretical results show that smaller microcompartments considerably enhance TcRR because the synthesized molecules, such as RNA and replicases, are more concentrated in smaller liposomes.
authors.affiliation: Institute for Academic Initiatives, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan. sunami-takeshi@ist.osaka-u.ac.jp.|Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Agency (JST), 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan. sunami-takeshi@ist.osaka-u.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Sunami
authors.affiliation: Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Agency (JST), 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.|Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: Norikazu
authors.initials: N
authors.lastname: Ichihashi
authors.affiliation: Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Agency (JST), 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: Takehiro
authors.initials: T
authors.lastname: Nishikawa
authors.affiliation: Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Agency (JST), 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Kazuta
authors.affiliation: Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Agency (JST), 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan. tetsuyayomo@gmail.com.|Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan. tetsuyayomo@gmail.com.|Graduate School of Frontier Biosciences, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan. tetsuyayomo@gmail.com.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Yomo
mesh_terms: D002474:Cell-Free System; D019418:Evolution, Chemical; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D020719:Glucuronides / Q000737:chemistry; D008956:Models, Chemical; D010316:Particle Size; D011777:Q beta Replicase / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D053835:Unilamellar Liposomes / Q000737:chemistry*; D005466:Fluorocarbons
chemical_list: D005452:Fluoresceins; D005456:Fluorescent Dyes; D005966:Glucuronidase; D020719:Glucuronides; D011777:Q beta Replicase; D012313:RNA; D053835:Unilamellar Liposomes; C000622621:5-(pentafluorobenzoylamino)fluorescein diglucuronide; D005466:Fluorocarbons
keywords: RNA replication; cell-free translation; fluorescence-activated cell sorter; liposomes; synthetic biology