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. 2021 Mar 24;11:6769. doi: 10.1038/s41598-021-85366-4

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Delivery of CRISPR/Cas9 components using a single TRBO vector. (a) Vector construct TRBOCas9-RCgGFP with gGFP expressed through the (−) sense RNA. (b) Western blot analysis of the Cas9 protein delivered using the TRBOCas9-RCgGFP and compared to other delivery vectors as indicated. (c) BsgI restriction assay detecting gene edited resistant band (red arrow) by the functional Cas9 protein and sgGFP of TRBOCas9-RCgGFP. (d) Sequence analyses of gene edited resistant bands (red arrows) showed various nucleotide indels at the BsgI target site.