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. 2019 Nov 25;9:17449. doi: 10.1038/s41598-019-53868-x

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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CRISPR/Cas9 mediated miR-29b knockdown in NIH/3T3 and HeLa cells. (a) The gRNAs were designed by submitting the whole length of hsa-mir-29b-1, mmu-mir-29b-1, mmu-mir-29b-2 onto gRNA design website http://crispr.mit.edu. The gRNA quality scores were calculated based on the number of potential off-targets and the number of in genes off-targets. (b) The gRNAs h-cas1, m-cas1, m-cas2 and m-cas3 locations, sequences and their PAMs were shown on the whole length sequences of hsa-mir-29b-1, mmu-mir-29b-1, mmu-mir-29b-2, respectively. (c) The expression levels of miR-29b in HeLa cell clones detected by qRT-PCR. (d) The expression levels of miR-29b in NIH/3T3 cells detected by qRT-PCR. Data shown represent the mean values of three independent experiments. Statistical analysis was performed using paired Student’s t-tests. *p < 0.05, ***p < 0.001, ****p < 0.0001.