Osteoarthritis and Cartilage CRISPR/CAS9 mediated genome engineering of human mesenchymal stem cells  G. van den Akker, H. van Beuningen, E. Blaney Davidson, P. van der Kraan  Osteoarthritis and Cartilage  Volume 24, (April 2016) DOI: 10.1016/j.joca.2016.01.445 Copyright © 2016 Terms and Conditions

Figure 1 CRISPR/Cas9 targetting of R-SMAD genes. A) Targetting strategy of exon 2 of R-SMAD genes with two different guide RNAs. By combing two guide RNAs the on-target efficiency can be improved. The expected band pattern after Surveyor nuclease treatment of a genomic PCR amplicon are indicated. B) Surveyor nuclease assay of an R-SMAD PCR amplicon in MSC. Numbers indicate CRISPR specific cleavage bands, the smallest 100 base pair band (4) is not visible on the agarose gel. The wild type amplicon (upper band) is strongly decreased upon addition of more CRISPR/Cas9 lentivirus. C) The double targetting strategy leads to an efficient decrease in protein expression in HEK293 cells as determined by immunoblotting. Osteoarthritis and Cartilage 2016 24, DOI: (10.1016/j.joca.2016.01.445) Copyright © 2016 Terms and Conditions