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. 2024 Jun 5;25:568. doi: 10.1186/s12864-024-10250-0

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© The Author(s) 2024

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Schematic diagram of the insertion of a conditional expression cassette using the modified i-PITT method. Primers are shown in red (see Supplemental Table 1). The region shown as “STOP” consists of three SV40-pA sequences. F14, F15, FRT-L, FRT-R: mutant FRT sites. SA: splice acceptor. WPRE: woodchuck hepatitis virus posttranscriptional regulatory element. Lox2272: mutant LoxP. bGH-pA: bovine growth hormone polyadenylationsignal sequence. SV40-pA: Simian virus 40 polyadenylation signal sequence. GOI: gene of interest.CAG: hybrid construct consisting of the cytomegalovirus enhancer fused to the chicken beta-actin promoter. attB, attP, attR, attL: attachment sites for PhiC31 integrase. GTNOP: a cassette containing “eGFP-T2A-Neomycin resistant gene-hOCT4-PolyA” [15]