1. Volume 8, Issue 1, Pages 90-98 (July 2003)
Comparison of gene expression after intraperitoneal delivery of AAV2 or AAV5 in utero 
Gerald S Lipshutz, Deborah Titre, Mary Brindle, Angelina R Bisconte, Christopher H Contag, Karin M.L Gaensler 
Molecular Therapy 
Volume 8, Issue 1, Pages (July 2003)
DOI: /S (03)
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

2. FIG. 1
Schematic representations of viral vectors constructed. AAV2 and AAV5 vectors were constructed using either the cytomegalovirus (CMV) promoter or the elongation factor 1 alpha (EF1α) promoter. AAV2 and AAV5 EF1α-luc contained the bovine growth hormone polyadenylation signal (BGH An), while the other two vectors included the SV40 polyadenylation sequences (SV40 An). The woodchuck posttranscriptional regulatory element (WPRE) was inserted 3′ of the luciferase cassette in the AAV5 vector. ITR, inverted terminal repeats.
Molecular Therapy 2003 8, 90-98DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

3. FIG. 2
Expression in 293 human embryonic kidney cells. Cells were transduced in vitro with 1 × 1010 genomes of rAAV, and luciferase expression was analyzed by luminometry. The y axis indicates the relative light units (RLU) per 1 × cells. The AAV vectors used, as well as the control, are designated above each bar. Error bars represent the standard error of the mean (SEM).
Molecular Therapy 2003 8, 90-98DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

4. FIG. 3
Graphic representation of longitudinal in vivo bioluminescence studies after AAV2 delivery. Mice were imaged repetitively over 18 months after in utero injection of rAAV2 containing either the CMV or the EF1α promoter. Total bioluminescence was quantified and plotted over time (indicated on the x axis). Data are plotted as counts per 5 min (indicated on the y axis). Each time point represents data from ≥4 animals.
Molecular Therapy 2003 8, 90-98DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

5. FIG. 4
Graphic representation of longitudinal in vivo bioluminescence studies after AAV5 delivery. Mice were serially followed over 15 months after in utero injection of rAAV5 containing the CMV promoter. Total bioluminescence was quantified and plotted over time (indicated on the x axis). Data are plotted as counts per 1 s (indicated on the y axis). This imaging system is more sensitive than equipment used in Fig. 3, as the data were collected over 1 s. Each time point represents data from ≥4 animals.
Molecular Therapy 2003 8, 90-98DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

6. FIG. 5
Longitudinal in vivo bioluminescence imaging after delivery of AAV5 CMV-luc-WPRE. Serial images captured from 2 weeks after birth to 15 months of life in analysis of a mouse injected in utero with 3 × 1011 genomes of AAV5 CMV-luc-WPRE. Image analysis was performed with the same parameters at each time point. The age of the animal at each time point is indicated above the image. The reference pseudocolor scale is the same at all time points.
Molecular Therapy 2003 8, 90-98DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

7. FIG. 6
Graphic summary of in vivo bioluminescence imaging analysis of adult animals after in utero injection. Mice were transduced in utero with equal numbers of genomes of rAAV. Expression was quantified by bioluminescence imaging. The y axis indicates the counts per second in tissues transduced by each vector (n at each ≥5). The AAV vectors administered and the uninjected control are indicated above each bar. Error bars represent the SEM.
Molecular Therapy 2003 8, 90-98DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

8. FIG. 7
Analysis of tissue distribution of AAV2 and AAV5. (A) Graphic summary comparing the tissue expression of AAV2 and AAV5 CMV-promoter vectors after in utero intraperitoneal injection. The y axis indicates the relative light units (RLU)/μg protein, a measurement of the quantity of luciferase. On the x axis is the name of each tissue. (B) PCR summary of comparison of vector distribution of AAV5 CMV, (C) of AAV2 CMV, and (D) of negative control: β-actin was used as a control for genomic DNA loading. The gels are representative of all the mice examined.
Molecular Therapy 2003 8, 90-98DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions