single RNA translation dynamics in living cells.
Science 352(6292):1425–1429. https://doi.
org/10.1126/science.aaf0899
54. Pichon X, Bastide A, Safieddine A, Chouaib R,
Samacoits A, Basyuk E, Peter M, Mueller F,
Bertrand E (2016) Visualization of single
endogenous polysomes reveals the dynamics
of translation in live human cells. J Cell Biol
214(6):769–781. https://doi.org/10.1083/
jcb.201605024
55. Wu B, Eliscovich C, Yoon YJ, Singer RH
(2016) Translation dynamics of single
mRNAs in live cells and neurons. Science 352
(6292):1430–1435.
https://doi.org/10.
1126/science.aaf1084
56. Garcia JF, Parker R (2015) MS2 coat proteins
bound to yeast mRNAs block 5
0 to 3
0 degradation and trap mRNA decay products: implications for the localization of mRNAs by
MS2-MCP system. RNA 21(8):1393–1395.
https://doi.org/10.1261/rna.051797.115
57. Haimovich G, Zabezhinsky D, Haas B,
Slobodin B, Purushothaman P, Fan L, Levin
JZ, Nusbaum C, Gerst JE (2016) Use of the
MS2 aptamer and coat protein for RNA localization in yeast: A response to “MS2 coat proteins bound to yeast mRNAs block 5
0 to 3
0
degradation and trap mRNA decay products:
implications for the localization of mRNAs by
MS2-MCP system”. RNA 22(5):660–666.
https://doi.org/10.1261/rna.055095.115
58. Tutucci E, Vera M, Biswas J, Garcia J, Parker R,
Singer RH (2018) An improved MS2 system
for accurate reporting of the mRNA life cycle.
Nat Methods 15(1):81–89. https://doi.org/
10.1038/nmeth.4502
59. Heinrich S, Sidler CL, Azzalin CM, Weis K
(2017) Stem-loop RNA labeling can affect
nuclear and cytoplasmic mRNA processing.
RNA 23(2):134–141. https://doi.org/10.
1261/rna.057786.116
60. Tantale K, Mueller F, Kozulic-Pirher A,
Lesne A, Victor JM, Robert MC, Capozi S,
Chouaib R, Backer V, Mateos-Langerak J,
Darzacq X, Zimmer C, Basyuk E, Bertrand E
(2016) A single-molecule view of transcription
reveals convoys of RNA polymerases and multiscale bursting. Nat Commun 7:12248.
https://doi.org/10.1038/ncomms12248
61. Chao JA, Patskovsky Y, Almo SC, Singer RH
(2008) Structural basis for the coevolution of a
viral RNA-protein complex. Nat Struct Mol
Biol 15(1):103–105. https://doi.org/10.
1038/nsmb1327
62. Larson DR, Zenklusen D, Wu B, Chao JA,
Singer RH (2011) Real-time observation of
transcription initiation and elongation on an
endogenous yeast gene. Science 332
(6028):475–478. https://doi.org/10.1126/
science.1202142
63. Saroufim MA, Bensidoun P, Raymond P,
Rahman S, Krause MR, Oeffinger M, Zenklusen D (2015) The nuclear basket mediates perinuclear mRNA scanning in budding yeast. J
Cell Biol 211(6):1131–1140. https://doi.
org/10.1083/jcb.201503070
64. Wang C, Han B, Zhou R, Zhuang X (2016)
Real-time imaging of translation on single
mrna transcripts in live cells. Cell 165
(4):990–1001. https://doi.org/10.1016/j.
cell.2016.04.040
65. Yan X, Hoek TA, Vale RD, Tanenbaum ME
(2016) Dynamics of translation of single
mRNA molecules In vivo. Cell 165
(4):976–989. https://doi.org/10.1016/j.cell.
2016.04.034
66. Das S, Moon HC, Singer RH, Park HY (2018)
A transgenic mouse for imaging activitydependent dynamics of endogenous Arc
mRNA in live neurons. Sci Adv 4(6):
eaar3448. https://doi.org/10.1126/sciadv.
aar3448
67. Chen J, Nikolaitchik O, Singh J, Wright A,
Bencsics CE, Coffin JM, Ni N, Lockett S,
Pathak VK, Hu WS (2009) High efficiency of
HIV-1 genomic RNA packaging and heterozygote formation revealed by single virion analysis. Proc Natl Acad Sci U S A 106
(32):13535–13540.
https://doi.org/10.
1073/pnas.0906822106
68. Lange S, Katayama Y, Schmid M, Burkacky O,
Brauchle C, Lamb DC, Jansen RP (2008)
Simultaneous transport of different localized
mRNA species revealed by live-cell imaging.
Traffic 9(8):1256–1267. https://doi.org/10.
1111/j.1600-0854.2008.00763.x
69. Brodsky AS, Silver PA (2002) Identifying proteins that affect mRNA localization in living
cells. Methods 26(2):151–155. https://doi.
org/10.1016/S1046-2023(02)00017-8
70. Takizawa PA, Vale RD (2000) The myosin
motor, Myo4p, binds Ash1 mRNA via the
adapter protein, She3p. Proc Natl Acad Sci U
S A 97(10):5273–5278. https://doi.org/10.
1073/pnas.080585897
71. Rodriguez EA, Campbell RE, Lin JY, Lin MZ,
Miyawaki A, Palmer AE, Shu X, Zhang J, Tsien
RY (2017) The growing and glowing toolbox
of fluorescent and photoactive proteins. Trends
Biochem Sci 42(2):111–129. https://doi.org/
10.1016/j.tibs.2016.09.010
72. Specht EA, Braselmann E, Palmer AE (2017) A
critical and comparative review of fluorescent
tools for live-cell imaging. Annu Rev Physiol
Imaging Single mRNAs in Living Eukaryotic Cells
143
Science 352(6292):1425–1429. https://doi.
org/10.1126/science.aaf0899
54. Pichon X, Bastide A, Safieddine A, Chouaib R,
Samacoits A, Basyuk E, Peter M, Mueller F,
Bertrand E (2016) Visualization of single
endogenous polysomes reveals the dynamics
of translation in live human cells. J Cell Biol
214(6):769–781. https://doi.org/10.1083/
jcb.201605024
55. Wu B, Eliscovich C, Yoon YJ, Singer RH
(2016) Translation dynamics of single
mRNAs in live cells and neurons. Science 352
(6292):1430–1435.
https://doi.org/10.
1126/science.aaf1084
56. Garcia JF, Parker R (2015) MS2 coat proteins
bound to yeast mRNAs block 5
0 to 3
0 degradation and trap mRNA decay products: implications for the localization of mRNAs by
MS2-MCP system. RNA 21(8):1393–1395.
https://doi.org/10.1261/rna.051797.115
57. Haimovich G, Zabezhinsky D, Haas B,
Slobodin B, Purushothaman P, Fan L, Levin
JZ, Nusbaum C, Gerst JE (2016) Use of the
MS2 aptamer and coat protein for RNA localization in yeast: A response to “MS2 coat proteins bound to yeast mRNAs block 5
0 to 3
0
degradation and trap mRNA decay products:
implications for the localization of mRNAs by
MS2-MCP system”. RNA 22(5):660–666.
https://doi.org/10.1261/rna.055095.115
58. Tutucci E, Vera M, Biswas J, Garcia J, Parker R,
Singer RH (2018) An improved MS2 system
for accurate reporting of the mRNA life cycle.
Nat Methods 15(1):81–89. https://doi.org/
10.1038/nmeth.4502
59. Heinrich S, Sidler CL, Azzalin CM, Weis K
(2017) Stem-loop RNA labeling can affect
nuclear and cytoplasmic mRNA processing.
RNA 23(2):134–141. https://doi.org/10.
1261/rna.057786.116
60. Tantale K, Mueller F, Kozulic-Pirher A,
Lesne A, Victor JM, Robert MC, Capozi S,
Chouaib R, Backer V, Mateos-Langerak J,
Darzacq X, Zimmer C, Basyuk E, Bertrand E
(2016) A single-molecule view of transcription
reveals convoys of RNA polymerases and multiscale bursting. Nat Commun 7:12248.
https://doi.org/10.1038/ncomms12248
61. Chao JA, Patskovsky Y, Almo SC, Singer RH
(2008) Structural basis for the coevolution of a
viral RNA-protein complex. Nat Struct Mol
Biol 15(1):103–105. https://doi.org/10.
1038/nsmb1327
62. Larson DR, Zenklusen D, Wu B, Chao JA,
Singer RH (2011) Real-time observation of
transcription initiation and elongation on an
endogenous yeast gene. Science 332
(6028):475–478. https://doi.org/10.1126/
science.1202142
63. Saroufim MA, Bensidoun P, Raymond P,
Rahman S, Krause MR, Oeffinger M, Zenklusen D (2015) The nuclear basket mediates perinuclear mRNA scanning in budding yeast. J
Cell Biol 211(6):1131–1140. https://doi.
org/10.1083/jcb.201503070
64. Wang C, Han B, Zhou R, Zhuang X (2016)
Real-time imaging of translation on single
mrna transcripts in live cells. Cell 165
(4):990–1001. https://doi.org/10.1016/j.
cell.2016.04.040
65. Yan X, Hoek TA, Vale RD, Tanenbaum ME
(2016) Dynamics of translation of single
mRNA molecules In vivo. Cell 165
(4):976–989. https://doi.org/10.1016/j.cell.
2016.04.034
66. Das S, Moon HC, Singer RH, Park HY (2018)
A transgenic mouse for imaging activitydependent dynamics of endogenous Arc
mRNA in live neurons. Sci Adv 4(6):
eaar3448. https://doi.org/10.1126/sciadv.
aar3448
67. Chen J, Nikolaitchik O, Singh J, Wright A,
Bencsics CE, Coffin JM, Ni N, Lockett S,
Pathak VK, Hu WS (2009) High efficiency of
HIV-1 genomic RNA packaging and heterozygote formation revealed by single virion analysis. Proc Natl Acad Sci U S A 106
(32):13535–13540.
https://doi.org/10.
1073/pnas.0906822106
68. Lange S, Katayama Y, Schmid M, Burkacky O,
Brauchle C, Lamb DC, Jansen RP (2008)
Simultaneous transport of different localized
mRNA species revealed by live-cell imaging.
Traffic 9(8):1256–1267. https://doi.org/10.
1111/j.1600-0854.2008.00763.x
69. Brodsky AS, Silver PA (2002) Identifying proteins that affect mRNA localization in living
cells. Methods 26(2):151–155. https://doi.
org/10.1016/S1046-2023(02)00017-8
70. Takizawa PA, Vale RD (2000) The myosin
motor, Myo4p, binds Ash1 mRNA via the
adapter protein, She3p. Proc Natl Acad Sci U
S A 97(10):5273–5278. https://doi.org/10.
1073/pnas.080585897
71. Rodriguez EA, Campbell RE, Lin JY, Lin MZ,
Miyawaki A, Palmer AE, Shu X, Zhang J, Tsien
RY (2017) The growing and glowing toolbox
of fluorescent and photoactive proteins. Trends
Biochem Sci 42(2):111–129. https://doi.org/
10.1016/j.tibs.2016.09.010
72. Specht EA, Braselmann E, Palmer AE (2017) A
critical and comparative review of fluorescent
tools for live-cell imaging. Annu Rev Physiol
Imaging Single mRNAs in Living Eukaryotic Cells
143