The DOI matches its Crossref record; 10 of the 10 references it cites were found in a registry or an index.
Targeting non-coding RNAs with the CRISPR/Cas9 system in human cell lines
Open accessAbstract
The CRISPR/Cas has been recently shown to be a powerful genome-editing tool in a variety of organisms. However, these studies are mainly focused on protein-coding genes. The present study aims to determine whether this technology can be applied to non-coding genes. One of the challenges for knockout of non-coding genes is that a small deletion or insertion generated by the standard CRISPR/Cas system may not necessarily lead to functional loss of a given non-coding gene because of lacking an open reading frame, especially in polyploidy human cell lines. To overcome this challenge, we adopt a selection system that allows for marker genes to integrate into the genome through homologous recombination (HR). Moreover, we construct a dual guide RNA vector that can make two cuts simultaneously at designated sites such that a large fragment can be deleted. With these approaches, we are able to successfully generate knockouts for miR-21, miR-29a, lncRNA-21A, UCA1 and AK023948 in various human cell lines. Finally, we show that the HR-mediated targeting efficiency can be further improved by suppression of the non-homologous end joining pathway. Together, these results demonstrate the feasibility of knockout for non-coding genes by the CRISPR/Cas system in human cell lines.
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Cites
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J. A., & Charpentier, E. (2012) · Science · doi:10.1126/science.1225829 · Cited by 17,116
Multiplex Genome Engineering Using CRISPR/Cas Systems
Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W., Marraffini, L. A., et al. (2013) · Science · doi:10.1126/science.1231143 · Cited by 15,339
RNA-Guided Human Genome Engineering via Cas9
Mali, P., Yang, L., Esvelt, K. M., Aach, J., Guell, M., DiCarlo, J. E., Norville, J. E., & Church, G. M. (2013) · Science · doi:10.1126/science.1232033 · Cited by 9,210
Wang, H., Yang, H., Shivalila, C. S., Dawlaty, M. M., Cheng, A. W., Zhang, F., & Jaenisch, R. (2013) · Cell · doi:10.1016/j.cell.2013.04.025 · Cited by 3,495
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
RNA-guided editing of bacterial genomes using CRISPR-Cas systems
Jiang, W., Bikard, D., Cox, D., Zhang, F., & Marraffini, L. A. (2013) · Nature Biotechnology · doi:10.1038/nbt.2508 · Cited by 2,429
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Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease
Cho, S. W., Kim, S., Kim, J. M., & Kim, J.-S. (2013) · Nature Biotechnology · doi:10.1038/nbt.2507 · Cited by 1,863
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Self-citationLin, Y., Cradick, T. J., Brown, M. T., Deshmukh, H., Ranjan, P., Sarode, N., Wile, B. M., Vertino, P. M., Stewart, F. J., & Bao, G. (2014) · Nucleic Acids Research · doi:10.1093/nar/gku402 · Cited by 634 · cited 8×
“Therefore, our dual gRNA approach increases the targeting efficiency by CRISPR/Cas9 system, demonstrating its feasibility for targeting lncRNAs with ei- ther a short fragment (475 bp for lncRNA-21A) or a relatively long fragment (>5.6 kb for UCA1).”
BackgroundInfluential
Cited by
The emerging role of lncRNAs in cancer
Huarte, M. (2015) · Nature Medicine · doi:10.1038/nm.3981 · Cited by 2,702
Lu, T. X., & Rothenberg, M. E. (2018) · Journal of Allergy and Clinical Immunology · doi:10.1016/j.jaci.2017.08.034 · Cited by 2,425
CRISPRi-based genome-scale identification of functional long noncoding RNA loci in human cells
Liu, S. J., Horlbeck, M. A., Cho, S. W., Birk, H. S., Malatesta, M., He, D., Attenello, F. J., Villalta, J. E., Cho, M. Y., Chen, Y., et al. (2017) · Science · doi:10.1126/science.aah7111 · Cited by 746
MicroRNAs as regulatory elements in immune system logic
Mehta, A., & Baltimore, D. (2016) · Nature Reviews Immunology · doi:10.1038/nri.2016.40 · Cited by 704
Potential of long non‐coding RNAs in cancer patients: From biomarkers to therapeutic targets
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Metadata
- Authors
- Ho, T.-T., Zhou, N., Huang, J., Koirala, P., Xu, M., Fung, R., Wu, F., & Mo, Y.-Y.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2014-11-20
- Volume
- 43
- Issue
- 3
- Pages
- e17-e17
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2167171238
- Cited by
- 244
- References
- 10
- FWCI
- 11.32
- Type
- journal article
- Language
- en
- Open access
- gold
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