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. 2018 May 29;8:8214. doi: 10.1038/s41598-018-26439-9

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© The Author(s) 2018

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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Prkdc gene editing strategy and outcomes, ZFN design and gene expression, and cutting activity in scid fibroblasts. (a) Schematic of Prkdc, including location of scid and ZFN target sites, structure of ZFN and sequences of various Prkdc alleles around scid site. (b) Production of ZFN monomers from SFFV-driven IPLV and IDLV (qPCR MOI 100). Proteins were extracted 3 d after transduction. The blotting revealed FLAG-tagged ZFN monomers (42 and 38.5 kDa for ZFN1 & ZFN2, respectively), with α-tubulin used as a loading control (49 kDa). (c) Cel-I analysis of Prkdc cutting by ZFN. Scid fibroblasts were transduced with IPLV- and IDLV-ZFN at the indicated MOI and genomic DNA was extracted 3 d later. The Cel-I-digested PCR products were separated on a polyacrylamide gel and diagnostic bands (black arrows) were used to calculate the % of indels induced by the various ZFN treatments and indicated on the gel image.