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. 2019 Nov 21;9:17312. doi: 10.1038/s41598-019-53611-6

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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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Transgene design and gross phenotype of Tnfsf11 targeted CRISPRi mice. (a) Schematic representation of the transgene used to create Tnfsf11 targeted CRISPRi mice. The dCas9::KRAB is under control of the CAG promoter which consists of the cytomegalovirus early enhancer; the promoter, first exon, and first intron of the chicken beta-actin gene; and the splice acceptor of the rabbit beta-globin gene. The sgRNA scaffold containing the targeting sequence of sgRNA-3 from the in vitro experiments was placed under the control of the mouse U6 small RNA promoter in the opposite direction of the CAG-driven dCas9::KRAB. (b) The top and middle images of each column are micro-CT generated images of the skull and right femur, respectively, of the indicated lines at five weeks of age. The images in the bottom row demonstrate the presence or lack of inguinal lymph nodes in each line. The dashed circle indicates the area of tissue where inguinal lymph node development normally occurs.