Table 1.

Liver Analysis of Treated PKU Mice Sacrificed at Different Timepoints

Mouse and Treatment	Timepoint of Sacrification (Number of Mice)	PAH Enzyme Activity		Relative Pah-mRNA Gene Expression (fold in %)	DNA Copy Number per Diploid Genome
		(in mU/mg)	(in % of wt)
Wild-type, untreated	(n = 3)	55.6 ± 10.6	100	n.d.	n.a.
PKU, untreated	(n = 3)	< 0.2	< 0.3	100	n.a.
PKU + MC (77 μg)	day 4 (n = 2)	37.4 ± 25.8	67.1 ± 46.4	215 ± 24	1345 ± 1153
	day 10 (n = 3)	9.0 ± 2.4	16.1 ± 4.4	217 ± 64	119 ± 62
	day 52 (n = 1)	8.1	14.6	199	16
	day 179 (n = 2)	5.2 ± 4.4	9.3 ± 7.9	131 ± 13	13 ± 7
PKU 1 MC (7.7 μg)	day 10 (n = 1)	1.9	3.4	112	8
PKU 1 MC (0.77 μg)	day 10 (n = 1)	< 0.2	< 0.2	141	0.2
PKU 1 PP (219 μg)	day 4 (n = 2)	< 0.2	< 0.2	93 ± 28	285 ± 92
	day 10 (n = 3)	0.6 ± 0.1	1.0 ± 0.2	151 ± 17	142 ± 27
	day 52 (n = 2)	< 0.2	< 0.2	123 ± 19	13 ± 7
PKU 1 MC & Stuffer* (219 μg)	day 14 (n = 2)	1.4 ± 0.9	2.4 ± 1.7	123 ± 8	27 ± 9

PAH enzyme activity, Pah-mRNA gene expression and copy numbers of vector genomes in whole liver extracts of PKU (Pah^enu2) mice injected with parental plasmid (PP) pMC.PKU20, with minicircle (MC) MC.PKU20, or with MC plus stuffer DNA (pMC.BESPX, empty vector). For blood Phe level values over time see also Figure 2. The amount of vector injected, time point of sacrification and the numbers of mice analyzed are indicated. Note that 219 μg of PP, or 77 μg of MC, is equivalent to 3 × 10¹³ DNA particles. n.d., not determined; n.a., not applicable.

^*Sum of MC.PKU20 (77 μg) plus stuffer DNA (142 μg; pMC.BESPX, empty vector); blood Phe levels dropped from 2150 ± 109 μmol/L (before injection) to 768 ± 161 μmol/L at day 5 postinjection, and to 536 ± 231 μmol/L at day 14 postinjection.