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. 2019 May 17;12:3849–3858. doi: 10.2147/OTT.S195661

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© 2019 Yang et al. This work is published and licensed by Dove Medical Press Limited

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Design of lentivirus, transfection, screening, and PCR sequencing of single-knockout (Sg-CXCR4 and Sg-CXCR7) and co-knockout (Sg-CXCR4+7) cell clones and verification of the protein expression level by Western blotting. (A and B) The knockout target sequence of CXCR4 is located in its exon 2, while the knockout target sequence of CXCR7 is located in exon 1. (C) In MDA-MB-231 cells, after transfection with Cas9/SgRNA lentivirus, the transfection efficiency was significant. (D–F) The PCR products of monoclonal cells after the knockout of CXCR4 and CXCR7 target genes were sequenced. (G) Western blotting was used to detect the expression of CXCR4 and CXCR7 in monoclonal cells.

Abbreviations: NC, negative control; ORIG, original; UNRE, unread; FRST, first; SCND, second; nt, nucleotide; MUTA, mutation.