targeted integration of transforming DNA fragments
[28]. The authors use this as an alternative approach to genetic
inhibition of the NHEJ pathway employing Δku70/Δku80
ortholog mutants of N. crassa which would require backcrossing for strain purification.
Acknowledgments
David Burnett and Alexander Lichius contributed equally to
this work.
References
1. Chin A, Le ´cuyer E (2017) RNA localization:
making its way to the center stage. BBA-Gen
Subjects 1861:2956–2970. https://doi.org/
10.1016/j.bbagen.2017.06.011
2. Ryder PV, Lerit DA (2018) RNA localization
regulates diverse and dynamic cellular processes. Traffic 19:496–502. https://doi.org/
10.1111/tra.12571
3. Suter B (2018) RNA localization and transport. Biochim Biophys Acta Gene Regul
Mech 1861:938–951. https://doi.org/10.
1016/j.bbagrm.2018.08.004
4. Tilsner J, Linnik O, Christensen NM, Bell K,
Roberts IM, Lacomme C, Oparka KJ (2009)
Live-cell imaging of viral RNA genomes using
a pumilio-based reporter. Plant J 57:758–770.
https://doi.org/10.1111/j.1365-313X.
2008.03720.x
5. Chitwood DH, Nogueira FTS, Howell MD,
Montgomery TA, Carrington JC, Timmermans MCP (2009) Pattern formation via
small RNA mobility. Genes Dev 23:549–554.
https://doi.org/10.1101/gad.1770009
6. Zarnack K, Feldbru ¨gge M (2007) mRNA trafficking in fungi. Mol Gen Genomics
278:347–359.
https://doi.org/10.1007/
s00438-007-0271-8
7. Ko ¨nig J, Baumann S, Koepke J, Pohlmann T,
Zarnack K, Feldbru ¨gge M (2009) The fungal
RNA-binding protein Rrm4 mediates longdistance transport of ubi1 and rho3 mRNAs.
EMBO J 28:1855–1866. https://doi.org/10.
1038/emboj.2009.145
8. Christensen NM, Oparka KJ, Tilsner J (2010)
Advances in imaging RNA in plants. Trends
Plant Sci 15:196–203. https://doi.org/10.
1016/j.tplants.2010.01.005
9. Chao JA, Lionnet T (2018) Imaging the life
and death of mRNAs in single cells. Cold
Spring Harb Perspect Biol 10:a032086.
https://doi.org/10.1101/cshperspect.
a032086
10. Cheong C-G, Tanaka Hall TM (2006) Engineering RNA sequence specificity of pumilio
repeats. Proc Natl Acad Sci U S A
103:13635–13639.
https://doi.org/10.
1073/pnas.0606294103
11. Cerny RE, Qi Y, Aydt CM, Huang S, Listello
JJ, Fabbri BJ, Conner TW, Crossland L, Huang
J (2003) RNA-binding protein-mediated
translational repression of transgene expression
in plants. Plant Mol Biol 52:357–369. https://
doi.org/10.1023/a:1023953130574
12. Heinrich S, Sidler CL, Azzalin CM, Weis K
(2017) Stem–loop RNA labeling can affect
nuclear and cytoplasmic mRNA processing.
RNA
23:134–141.
https://doi.org/10.
1261/rna.057786
13. Sternberg SH, Haurwitz RE, Doudna JA
(2012) Mechanism of substrate selection by a
highly specific CRISPR endoribonuclease.
RNA
18:661–672.
https://doi.org/10.
1261/rna.030882.111
14. Lee HY, Haurwitz RE, Apffel A, Zhou K,
Smart B, Wenger CD, Laderman S, Bruhn L,
Doudna JA (2013) RNA–protein analysis using
a conditional CRISPR nuclease. Proc Natl
Acad Sci U S A 110:5416–5421
15. Shukla S, Dickmeis C, Nagarajan AS, Fischer R,
Commandeur U, Steinmetz NF (2014) Molecular farming of fluorescent virus-based nanoparticles for optical imaging in plants, human
cells and mouse models. Biomater Sci
2:784–797.
https://doi.org/10.1039/
c3bm60277j
16. Haurwitz RE, Jinek M, Wiedenheft B, Zhou K,
Doudna JA (2010) Sequence- and structurespecific RNA processing by a CRISPR endonuclease. Science 329:1355–1358. https://doi.
org/10.1126/science.1192272
RNA Imaging with Csy4
177
[28]. The authors use this as an alternative approach to genetic
inhibition of the NHEJ pathway employing Δku70/Δku80
ortholog mutants of N. crassa which would require backcrossing for strain purification.
Acknowledgments
David Burnett and Alexander Lichius contributed equally to
this work.
References
1. Chin A, Le ´cuyer E (2017) RNA localization:
making its way to the center stage. BBA-Gen
Subjects 1861:2956–2970. https://doi.org/
10.1016/j.bbagen.2017.06.011
2. Ryder PV, Lerit DA (2018) RNA localization
regulates diverse and dynamic cellular processes. Traffic 19:496–502. https://doi.org/
10.1111/tra.12571
3. Suter B (2018) RNA localization and transport. Biochim Biophys Acta Gene Regul
Mech 1861:938–951. https://doi.org/10.
1016/j.bbagrm.2018.08.004
4. Tilsner J, Linnik O, Christensen NM, Bell K,
Roberts IM, Lacomme C, Oparka KJ (2009)
Live-cell imaging of viral RNA genomes using
a pumilio-based reporter. Plant J 57:758–770.
https://doi.org/10.1111/j.1365-313X.
2008.03720.x
5. Chitwood DH, Nogueira FTS, Howell MD,
Montgomery TA, Carrington JC, Timmermans MCP (2009) Pattern formation via
small RNA mobility. Genes Dev 23:549–554.
https://doi.org/10.1101/gad.1770009
6. Zarnack K, Feldbru ¨gge M (2007) mRNA trafficking in fungi. Mol Gen Genomics
278:347–359.
https://doi.org/10.1007/
s00438-007-0271-8
7. Ko ¨nig J, Baumann S, Koepke J, Pohlmann T,
Zarnack K, Feldbru ¨gge M (2009) The fungal
RNA-binding protein Rrm4 mediates longdistance transport of ubi1 and rho3 mRNAs.
EMBO J 28:1855–1866. https://doi.org/10.
1038/emboj.2009.145
8. Christensen NM, Oparka KJ, Tilsner J (2010)
Advances in imaging RNA in plants. Trends
Plant Sci 15:196–203. https://doi.org/10.
1016/j.tplants.2010.01.005
9. Chao JA, Lionnet T (2018) Imaging the life
and death of mRNAs in single cells. Cold
Spring Harb Perspect Biol 10:a032086.
https://doi.org/10.1101/cshperspect.
a032086
10. Cheong C-G, Tanaka Hall TM (2006) Engineering RNA sequence specificity of pumilio
repeats. Proc Natl Acad Sci U S A
103:13635–13639.
https://doi.org/10.
1073/pnas.0606294103
11. Cerny RE, Qi Y, Aydt CM, Huang S, Listello
JJ, Fabbri BJ, Conner TW, Crossland L, Huang
J (2003) RNA-binding protein-mediated
translational repression of transgene expression
in plants. Plant Mol Biol 52:357–369. https://
doi.org/10.1023/a:1023953130574
12. Heinrich S, Sidler CL, Azzalin CM, Weis K
(2017) Stem–loop RNA labeling can affect
nuclear and cytoplasmic mRNA processing.
RNA
23:134–141.
https://doi.org/10.
1261/rna.057786
13. Sternberg SH, Haurwitz RE, Doudna JA
(2012) Mechanism of substrate selection by a
highly specific CRISPR endoribonuclease.
RNA
18:661–672.
https://doi.org/10.
1261/rna.030882.111
14. Lee HY, Haurwitz RE, Apffel A, Zhou K,
Smart B, Wenger CD, Laderman S, Bruhn L,
Doudna JA (2013) RNA–protein analysis using
a conditional CRISPR nuclease. Proc Natl
Acad Sci U S A 110:5416–5421
15. Shukla S, Dickmeis C, Nagarajan AS, Fischer R,
Commandeur U, Steinmetz NF (2014) Molecular farming of fluorescent virus-based nanoparticles for optical imaging in plants, human
cells and mouse models. Biomater Sci
2:784–797.
https://doi.org/10.1039/
c3bm60277j
16. Haurwitz RE, Jinek M, Wiedenheft B, Zhou K,
Doudna JA (2010) Sequence- and structurespecific RNA processing by a CRISPR endonuclease. Science 329:1355–1358. https://doi.
org/10.1126/science.1192272
RNA Imaging with Csy4
177