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01.007
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org/10.1126/science.1179555
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science.1225829
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org/10.1038/nature13579
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https://doi.org/10.
1126/science.aab1452
9. Sternberg SH, Redding S, Jinek M, Greene
EC, Doudna JA (2014) DNA interrogation
by the CRISPR RNA-guided endonuclease
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org/10.1038/nature13011
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Habib N, Hsu PD, Wu X, Jiang W, Marraffini
LA, Zhang F (2013) Multiplex genome engineering using CRISPR/Cas systems. Science
339(6121):819–823.
https://doi.org/10.
1126/science.1231143
11. Mali P, Yang L, Esvelt KM, Aach J, Guell M,
DiCarlo JE, Norville JE, Church GM (2013)
RNA-guided human genome engineering via
Cas9. Science 339(6121):823–826. https://
doi.org/10.1126/science.1232033
12. Jinek M, East A, Cheng A, Lin S, Ma E,
Doudna J (2013) RNA-programmed genome
editing in human cells. eLife 2:e00471.
https://doi.org/10.7554/eLife.00471
13. Chen B, Gilbert LA, Cimini BA,
Schnitzbauer J, Zhang W, Li GW, Park J,
Blackburn EH, Weissman JS, Qi LS, Huang B
(2013) Dynamic imaging of genomic loci in
living human cells by an optimized CRISPR/
Cas system. Cell 155(7):1479–1491. https://
doi.org/10.1016/j.cell.2013.12.001
14. Shao S, Chang L, Sun Y, Hou Y, Fan X, Sun Y
(2017) Multiplexed sgRNA expression allows
versatile single nonrepetitive DNA labeling and
endogenous gene regulation. ACS Synth Biol.
https://doi.org/10.1021/acssynbio.7b00268
15. Ye H, Rong Z, Lin Y (2017) Live cell imaging
of genomic loci using dCas9-SunTag system
and a bright fluorescent protein. Protein Cell
8(11):853–855. https://doi.org/10.1007/
s13238-017-0460-0
16. Shao S, Zhang W, Hu H, Xue B, Qin J, Sun C,
Sun Y, Wei W, Sun Y (2016) Long-term dualcolor tracking of genomic loci by modified
sgRNAs of the CRISPR/Cas9 system. Nucleic
Acids Res 44(9):e86. https://doi.org/10.
1093/nar/gkw066
17. Fu Y, Rocha PP, Luo VM, Raviram R, Deng Y,
Mazzoni EO, Skok JA (2016) CRISPR-dCas9
and sgRNA scaffolds enable dual-colour live
imaging of satellite sequences and repeatenriched individual loci. Nat Commun
7:11707.
https://doi.org/10.1038/
ncomms11707
18. Qin P, Parlak M, Kuscu C, Bandaria J, Mir M,
Szlachta K, Singh R, Darzacq X, Yildiz A, Adli
M (2017) Live cell imaging of low- and
non-repetitive chromosome loci using
CRISPR-Cas9. Nat Commun 8:14725.
https://doi.org/10.1038/ncomms14725
19. Wang S, Su JH, Zhang F, Zhuang X (2016) An
RNA-aptamer-based two-color CRISPR labeling system. Sci Rep 6:26857. https://doi.org/
10.1038/srep26857
20. Ma H, Tu LC, Naseri A, Huisman M, Zhang S,
Grunwald D, Pederson T (2016) Multiplexed
labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Nat
Biotechnol 34(5):528–530. https://doi.org/
10.1038/nbt.3526
21. Gibcus JH, Samejima K, Goloborodko A,
Samejima I, Naumova N, Nuebler J, Kanemaki
MT, Xie L, Paulson JR, Earnshaw WC, Mirny
LA, Dekker J (2018) A pathway for mitotic
Visualizing Genomic Loci by CRISPR/MB
371
1. Misteli T (2013) The cell biology of genomes:
bringing the double helix to life. Cell 152
(6):1209–1212. https://doi.org/10.1016/j.
cell.2013.02.048
2. Misteli T (2007) Beyond the sequence: cellular
organization of genome function. Cell 128
(4):787–800. https://doi.org/10.1016/j.cell.
2007.01.028
3. Chen B, Guan J, Huang B (2016) Imaging
specific genomic DNA in living cells. Annu
Rev Biophys 45:1–23. https://doi.org/10.
1146/annurev-biophys-062215-010830
4. Li T, Meng YQ, Yao RY, Han H, Wu L, Zhou
YN, Li ZQ, Zhang YF, Fu LG (2019) The
associations between left-hand digit ratio
(2D:4D) and puberty characteristics among
Chinese girls. Early Hum Dev 130:22–26.
https://doi.org/10.1016/j.earlhumdev.2019.
01.007
5. Horvath P, Barrangou R (2010) CRISPR/Cas,
the immune system of bacteria and archaea.
Science 327(5962):167–170. https://doi.
org/10.1126/science.1179555
6. Jinek M, Chylinski K, Fonfara I, Hauer M,
Doudna JA, Charpentier E (2012) A programmable dual-RNA-guided DNA endonuclease
in adaptive bacterial immunity. Science 337
(6096):816–821. https://doi.org/10.1126/
science.1225829
7. Anders C, Niewoehner O, Duerst A, Jinek M
(2014) Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease. Nature 513(7519):569–573. https://doi.
org/10.1038/nature13579
8. Jiang F, Zhou K, Ma L, Gressel S, Doudna JA
(2015) A Cas9-guide RNA complex preorganized for target DNA recognition. Science 348
(6242):1477–1481.
https://doi.org/10.
1126/science.aab1452
9. Sternberg SH, Redding S, Jinek M, Greene
EC, Doudna JA (2014) DNA interrogation
by the CRISPR RNA-guided endonuclease
Cas9. Nature 507(7490):62–67. https://doi.
org/10.1038/nature13011
10. Cong L, Ran FA, Cox D, Lin S, Barretto R,
Habib N, Hsu PD, Wu X, Jiang W, Marraffini
LA, Zhang F (2013) Multiplex genome engineering using CRISPR/Cas systems. Science
339(6121):819–823.
https://doi.org/10.
1126/science.1231143
11. Mali P, Yang L, Esvelt KM, Aach J, Guell M,
DiCarlo JE, Norville JE, Church GM (2013)
RNA-guided human genome engineering via
Cas9. Science 339(6121):823–826. https://
doi.org/10.1126/science.1232033
12. Jinek M, East A, Cheng A, Lin S, Ma E,
Doudna J (2013) RNA-programmed genome
editing in human cells. eLife 2:e00471.
https://doi.org/10.7554/eLife.00471
13. Chen B, Gilbert LA, Cimini BA,
Schnitzbauer J, Zhang W, Li GW, Park J,
Blackburn EH, Weissman JS, Qi LS, Huang B
(2013) Dynamic imaging of genomic loci in
living human cells by an optimized CRISPR/
Cas system. Cell 155(7):1479–1491. https://
doi.org/10.1016/j.cell.2013.12.001
14. Shao S, Chang L, Sun Y, Hou Y, Fan X, Sun Y
(2017) Multiplexed sgRNA expression allows
versatile single nonrepetitive DNA labeling and
endogenous gene regulation. ACS Synth Biol.
https://doi.org/10.1021/acssynbio.7b00268
15. Ye H, Rong Z, Lin Y (2017) Live cell imaging
of genomic loci using dCas9-SunTag system
and a bright fluorescent protein. Protein Cell
8(11):853–855. https://doi.org/10.1007/
s13238-017-0460-0
16. Shao S, Zhang W, Hu H, Xue B, Qin J, Sun C,
Sun Y, Wei W, Sun Y (2016) Long-term dualcolor tracking of genomic loci by modified
sgRNAs of the CRISPR/Cas9 system. Nucleic
Acids Res 44(9):e86. https://doi.org/10.
1093/nar/gkw066
17. Fu Y, Rocha PP, Luo VM, Raviram R, Deng Y,
Mazzoni EO, Skok JA (2016) CRISPR-dCas9
and sgRNA scaffolds enable dual-colour live
imaging of satellite sequences and repeatenriched individual loci. Nat Commun
7:11707.
https://doi.org/10.1038/
ncomms11707
18. Qin P, Parlak M, Kuscu C, Bandaria J, Mir M,
Szlachta K, Singh R, Darzacq X, Yildiz A, Adli
M (2017) Live cell imaging of low- and
non-repetitive chromosome loci using
CRISPR-Cas9. Nat Commun 8:14725.
https://doi.org/10.1038/ncomms14725
19. Wang S, Su JH, Zhang F, Zhuang X (2016) An
RNA-aptamer-based two-color CRISPR labeling system. Sci Rep 6:26857. https://doi.org/
10.1038/srep26857
20. Ma H, Tu LC, Naseri A, Huisman M, Zhang S,
Grunwald D, Pederson T (2016) Multiplexed
labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Nat
Biotechnol 34(5):528–530. https://doi.org/
10.1038/nbt.3526
21. Gibcus JH, Samejima K, Goloborodko A,
Samejima I, Naumova N, Nuebler J, Kanemaki
MT, Xie L, Paulson JR, Earnshaw WC, Mirny
LA, Dekker J (2018) A pathway for mitotic
Visualizing Genomic Loci by CRISPR/MB
371