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. 2020 Oct 15;10:17479. doi: 10.1038/s41598-020-74474-2

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© The Author(s) 2020

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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Flp-TAL recombinase can delete large genome DNA fragments. (A) Schematic of the deletion assay. Upon expression of FV71-TAL in cells, in which pTarget was pre-integrated into FL-61 (Fig. 4A), the DNA fragment located between two FL-71 sequences can be deleted (this DNA fragment contains ~ 4.7 kb of reporter DNA and ~ 10 kb of genome DNA). (B) PCR analysis of the FV71-TAL treated cells. The deletion activity was analyzed in three biological replicates (see Supplemental Fig. 4). FL-61(LJ) and FL-71(RJ), the PCR analysis of the respective left and right vector-genomic junctions of the integrated pTarget before and after the treatment with FV71-TAL. M, DNA ladder (NEB 2-log). (C) Sequencing of the deletion-specific PCR product FL-71(RJ) confirmed its identity.