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. 2023 Jul 13;24(14):11421. doi: 10.3390/ijms241411421

Figure 3.

Figure 3

Quantification of HR-dependent reconstitution efficiency of qDys and concatemerization events by ddPCR. (A) Schematic representation of the protocol used to calculate HR efficiency using specific primers for VCN analysis of the single and recombined vectors. (B) Cloud separation showing single and double positive droplets according to the dye used (VIC or FAM) and the probe concentration (1× or 2×). (C) Primer design strategy and protocol to quantify all the ITR-dependent concatemerization events. (D) Viral copy number (VCN) of 5′or 3′ vectors and the recombined qDys vector, performed on TA, GA, and DIA of mdx systemically injected with dual vector AAVs. (E) Homologous recombination (HR) and concatemerization efficiency, calculated as described in panel A and C and expressed as a percentage. (F,G) VCN analysis of qDys (F) and HR efficiency (G) in Young and Old mdx mice co-injected with 5′ and 3′ AAV vectors. Data are presented as mean ± SEM (n = 5) (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). ns, not significant.