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. 2017 Dec 6;32(4):1048–1051. doi: 10.1038/leu.2017.346

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© The Author(s) 2018, corrected publication 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 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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CRISPR/Cas9 allows for the fast and efficient generation of SPARC deficient NSG mice. (a) RFLP (PleI digest) and T7 endonuclease analysis of day 4 NSG embryos microinjected with gRNA#1, Cas9 and ssDNA template. Asterisks mark productively edited embryos. (b) Schematic view of the workflow to generate NSG-edited line (top) and RFLP analysis of tail DNA from chimeric F0 mice (bottom). Asterisks mark HDR-edited mice. A subset of F0 mice (red squares) were backcrossed to WT NSG mice to obtain N1 progeny and evaluate germline transmission of the HDR+ allele. (c) Western blot on bone marrow cells shows complete loss of SPARC protein in SPARC^HDR/HDR animals, whereas a significant reduction is also observed in heterozygous SPARC^HDR/+ mice. Sam68 is used as loading control.