Biochemical and Biophysical Research Communications
A simple and rapid method to determine the zygosity of uPA-transgenic SCID mice
Section snippets
Materials and methods
Breeding of the uPA-SCID mice. B6SJL-TgN(Alb1Plau)144Bri mice were back-crossed on CBySmn.CB17-Prkdcscid mice. Both strains were purchased from the Jackson Laboratories (Bar Harbor, Maine, USA). Screening for the SCID background was done with an in-house mouse IgM sandwich Elisa. Screening for the uPA transgene was done by PCR, initially using the genotyping protocol described by the Jackson Laboratories [10] and subsequently by the in-house method described herein. All mice were bred and
Sequencing of plasmid p3371C
Rhim et al. [2] noted that the insertion of the uPA transgene into the mouse genome resulted in the deletion of endogenous DNA at the integration site. These investigators picked up part of this sequence in plasmid p3371C (Fig. 1) and used the BamHI-fragment of this region as a probe for detection via Southern blotting.
The sequence of the deleted region was unknown but part of the plasmid has been sequenced. Starting from this known sequence we performed serial overlapping sequence reactions
Discussion
The need for an affordable, reliable, and ethically acceptable in vivo model to study HBV and HCV infections is high. The uPA-mouse model first described in 1990 represents one of the most promising tools to create this research tool. uPA-transgenic mice bred on an immune incompetent background like SCID, SCID/beige or RAG−/− allow a repopulation of the sick liver by allogeneic and xenogeneic hepatocytes. When transplanted with human hepatocytes these animals become susceptible to infections
Acknowledgements
The authors wish to thank Dr. R. Palmiter, Howard Hughes Medical Institute, University of Washington, for his kind donation of plasmid p3371C.
References (10)
- et al.
Neonatal bleeding in transgenic mice expressing urokinase-type plasminogen activator
Cell
(1990) - et al.
Hepatocyte transplantation into diseased mouse liver. Kinetics of parenchymal repopulation and identification of the proliferative capacity of tetraploid and octaploid hepatocytes
Am. J. Pathol.
(2000) - et al.
Repopulation of mouse liver with human hepatocytes and in vivo infection with hepatitis B virus
Hepatology
(2001) - et al.
Complete hepatic regeneration after somatic deletion of an albumin–plasminogen activator transgene
Cell
(1991) - et al.
Hepatitis C virus infection
N. Engl. J. Med.
(2001)