Fig. 3 | Nature Communications

Fig. 3

From: Genome-wide microhomologies enable precise template-free editing of biologically relevant deletion mutations

Fig. 3

@Target variant associated with muscular dystrophy created by MMEJ recapitulates DYSFERLIN loss-of-function. a Sequence verification of a precise 5 bp deletion mutation in DYSF. Deletion (dotted line), DSB location (pink bolt), µH (green), SpCas9 PAM (underline). b Altered protein sequence of DYSFERLIN caused by the 5 bp deletion mutation. c Schematic of the experimental procedure to induce myogenic differentiation in hiPSCs by overexpression of MYOD from a piggyBac transposon. Differentiation day (D). d Immunostaining for DYSFERLIN and MHC in differentiated hiPSC populations. Comparison of the isogenic parental cell line, three derived clones carrying the disease mutation and a muscular dystrophy patient derived hiPSC cell line. Scale bar indicates 100 µm; ratio of mCherry + cells measured by FACS in corresponding electroporated (EP) hiPSC populations indicated on the right

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