The DOI matches its Crossref record; 9 of the 9 references it cites were found in a registry or an index.
Rapid reverse genetic screening using CRISPR in zebrafish
Open accessAbstract
Identifying genes involved in biological processes is critical for understanding the molecular building blocks of life. We used engineered CRISPR (clustered regularly interspaced short palindromic repeats) to efficiently mutate specific loci in zebrafish (Danio rerio) and screen for genes involved in vertebrate biological processes. We found that increasing CRISPR efficiency by injecting optimized amounts of Cas9-encoding mRNA and multiplexing single guide RNAs (sgRNAs) allowed for phenocopy of known mutants across many phenotypes in embryos. We performed a proof-of-concept screen in which we used intersecting, multiplexed pool injections to examine 48 loci and identified two new genes involved in electrical-synapse formation. By deep sequencing target loci, we found that 90% of the genes were effectively screened. We conclude that CRISPR can be used as a powerful reverse genetic screening strategy in vivo in a vertebrate system.
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Cites
Fiji: an open-source platform for biological-image analysis
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A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
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Genome-scale CRISPR-Cas9 knockout screening in human cells
Shalem, O., Sanjana, N. E., Hartenian, E., Shi, X., Scott, D. A., Mikkelsen, T. S., Heckl, D., Ebert, B. L., Root, D. E., Doench, J. G., et al. (2014) · Science · doi:10.1126/science.1247005 · Cited by 5,234
The zebrafish reference genome sequence and its relationship to the human genome.
Howe, K., Clark, M. D., Torroja, C. F., Torrance, J., Berthelot, C., Muffato, M., Collins, J. E., Humphray, S., McLaren, K., Matthews, L., et al. (2013) · Nature · doi:10.1038/nature12111 · Cited by 5,100
DNA targeting specificity of RNA-guided Cas9 nucleases
Hsu, P. D., Scott, D. A., Weinstein, J. A., Ran, F. A., Konermann, S., Agarwala, V., Li, Y., Fine, E. J., Wu, X., Shalem, O., et al. (2013) · Nature Biotechnology · doi:10.1038/nbt.2647 · Cited by 4,741
Efficient genome editing in zebrafish using a CRISPR-Cas system
Hwang, W. Y., Fu, Y., Reyon, D., Maeder, M. L., Tsai, S. Q., Sander, J. D., Peterson, R. T., Yeh, J.-R. J., & Joung, J. K. (2013) · Nature Biotechnology · doi:10.1038/nbt.2501 · Cited by 2,937
Zebrafish trilobite identifies new roles for Strabismus in gastrulation and neuronal movements
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Cited by
CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo
Moreno-Mateos, M. A., Vejnar, C. E., Beaudoin, J.-D., Fernandez, J. P., Mis, E. K., Khokha, M. K., & Giraldez, A. J. (2015) · Nature Methods · doi:10.1038/nmeth.3543 · Cited by 1,290
Genome-Editing Technologies: Principles and Applications
Gaj, T., Sirk, S. J., Shui, S.-L., & Liu, J. (2016) · Cold Spring Harbor Perspectives in Biology · doi:10.1101/cshperspect.a023754 · Cited by 381
Neurons secrete miR-132-containing exosomes to regulate brain vascular integrity
Xu, B., Zhang, Y., Du, X.-F., Li, J., Zi, H.-X., Bu, J.-W., Yan, Y., Han, H., & Du, J.-L. (2017) · Cell Research · doi:10.1038/cr.2017.62 · Cited by 354
Applications of CRISPR–Cas systems in neuroscience
Heidenreich, M., & Zhang, F. (2016) · Nature Reviews Neuroscience · doi:10.1038/nrn.2015.2 · Cited by 283
Artificial Virus Delivers CRISPR-Cas9 System for Genome Editing of Cells in Mice
Li, L., Song, L., Liu, X., Yang, X., Li, X., He, T., Wang, N., Yang, S., Yu, C., Yin, T., et al. (2017) · ACS Nano · doi:10.1021/acsnano.6b04261 · Cited by 244
Metadata
- Authors
- Shah, A. N., Davey, C. F., Whitebirch, A. C., Miller, A. C., & Moens, C. B.
- Venue
- Nature Methods
- Published
- 2015-04-13
- Volume
- 12
- Issue
- 6
- Pages
- 535-540
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- OpenAlex
- W2093767301
- Cited by
- 351
- References
- 9
- FWCI
- 16.73
- Type
- journal article
- Language
- en
- Open access
- bronze
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