Liposome-mediated RNA transfection should be used with caution

被引:35
作者
Barreau, Carine
Dutertre, Stephanie
Paillard, Luc
Osborne, H. Beverley
机构
[1] Univ Rennes 1, UMR 6061, IFR 140, Fac Med,CNRS, F-35043 Rennes, France
[2] CNRS, UMR 6061, IFR 140, Fac Med, F-35043 Rennes, France
关键词
lipofection; AU-rich element; EDEN; mRNA degradation; localization;
D O I
10.1261/rna.191706
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
Liposome-mediated RNA transfection appears to present a number of advantages for studying the metabolism of reporter mRNAs in mammalian cells. This method is also widely used to transfect siRNAs. Here we describe results indicating that reporter mRNAs introduced into HeLa cells by liposomes do not present the expected behaviors. Namely, the stability of reporter mRNAs was independent of the presence or absence of an AUUUA instability element, a poly(A) tail, or even a 5' methylated cap. Confocal microscopy showed that fluorescent RNAs introduced by liposome-mediated transfection were present in discrete particles. These observations imply that a number of control experiments are required when using liposome to mediated RNA transfection, and the possible consequences are discussed.
引用
收藏
页码:1790 / 1793
页数:4
相关论文
共 11 条
  • [1] Postfertilization deadenylation of mRNAs in Xenopus laevis embryos is sufficient to cause their degradation at the blastula stage
    Audic, Y
    Omilli, F
    Osborne, HB
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 1997, 17 (01) : 209 - 218
  • [2] AU-RICH ELEMENTS - CHARACTERIZATION AND IMPORTANCE IN MESSENGER-RNA DEGRADATION
    CHEN, CYA
    SHYU, AB
    [J]. TRENDS IN BIOCHEMICAL SCIENCES, 1995, 20 (11) : 465 - 470
  • [3] EDEN-dependent translational repression of maternal mRNAs is conserved between Xenopus and Drosophila
    Ezzeddine, N
    Paillard, L
    Capri, M
    Maniey, D
    Bassez, T
    Aït-Ahmed, O
    Osborne, HB
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (01) : 257 - 262
  • [4] Stress granule assembly is mediated by prion-like aggregation of TIA-1
    Gilks, N
    Kedersha, N
    Ayodele, M
    Shen, L
    Stoecklin, G
    Dember, LM
    Anderson, P
    [J]. MOLECULAR BIOLOGY OF THE CELL, 2004, 15 (12) : 5383 - 5398
  • [5] MicroRNAs control translation initiation by inhibiting eukaryotic initiation factor 4E/cap and poly(A) tail function
    Humphreys, DT
    Westman, BJ
    Martin, DIK
    Preiss, T
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2005, 102 (47) : 16961 - 16966
  • [6] Stress granules: sites of mRNA triage that regulate mRNA stability and translatability
    Kedersha, N
    Anderson, P
    [J]. BIOCHEMICAL SOCIETY TRANSACTIONS, 2002, 30 : 963 - 969
  • [7] c-Jun ARE targets mRNA deadenylation by an EDEN-BP (embryo deadenylation element-binding protein)-dependent pathway
    Paillard, L
    Legagneux, V
    Maniey, D
    Osborne, HB
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (05) : 3232 - 3235
  • [8] Short RNAs repress translation after initiation in mammalian cells
    Petersen, CP
    Bordeleau, ME
    Pelletier, J
    Sharp, PA
    [J]. MOLECULAR CELL, 2006, 21 (04) : 533 - 542
  • [9] Inhibition of translational initiation by Let-7 microRNA in human cells
    Pillai, RS
    Bhattacharyya, SN
    Artus, CG
    Zoller, T
    Cougot, N
    Basyuk, E
    Bertrand, E
    Filipowicz, W
    [J]. SCIENCE, 2005, 309 (5740) : 1573 - 1576
  • [10] Metazoan oocyte and early embryo development program: a progression through translation regulatory cascades
    Vasudevan, S
    Seli, E
    Steitz, JA
    [J]. GENES & DEVELOPMENT, 2006, 20 (02) : 138 - 146