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. 2021 Apr 6;12:627050. doi: 10.3389/fgene.2021.627050

FIGURE 3.

FIGURE 3

Specificity and functionality of RP loop CRISPR/Cas9 complex. (A) Specificity of RP loop-CRSPR-/Cas9 was determined using the GeneArt® Genomic Cleavage Selection Kit that expressed orange fluorescent reporter, in 293K cells 72 h posttransfection, specific to the 11205G nucleotide of ND4. qPCR for mtDNA levels. (B) qRT-PCR for ND4 mRNA expression (C) was significantly lower in RP loop sgRNA-CRISPR/Cas9-transfected MEF cells, at 24 h posttransfection, compared with the MLS–Cas9 controls (n = 5, Student’s t-test, *p = 0.016; **p = 0.005).