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. 2021 Mar 16;11:54. doi: 10.1186/s13578-021-00566-9

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© The Author(s) 2021

Open AccessThis 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 — Agmo knockout-first allele construct and confirmation of cassette integration in the Agmo region of the mouse genome. a EUCOMM construct for Agmo knockout-first (Agmo-lacZ) allele. The critical exon 2 is flanked by loxP sites to create a floxed allele for conditional knockout. pA: polyadenylation sequence; FRT: FLP recognition target. b, c FISH experiments in heterozygous Agmo knockout (Agmo-lacZ tg/wt, b) and wild-type (WT, c) fibroblasts using BAC probes against chromosome 12 (red) and the cassette (green, red arrow). Insets show a 2x magnification of the region of interest. Fiber-FISH experiments show integration of the cassette (red signal, indicated by the red arrow) in the middle of the Agmo locus (green signal) in heterozygous cells (d). Wild-type fibers (e) showed no signal for the cassette