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dc.contributor.authorSonsteng, K. M.
dc.contributor.authorPrigge, Justin R.
dc.contributor.authorJune, Ronald K.
dc.contributor.authorSchmidt, Edward E.
dc.date.accessioned2015-02-20T19:40:46Z
dc.date.available2015-02-20T19:40:46Z
dc.date.issued2014-03
dc.identifier.citationSonsteng KM, Prigge JR, June RK, and Schmidt EE. Hydrodynamic Delivery of Cre Proteins to Lineage-Mark or Time-Stamp Hepatocytes In Situ. PLOS ONE, (In Press 2/2014).en_US
dc.identifier.issn1932-6203
dc.identifier.urihttps://scholarworks.montana.edu/xmlui/handle/1/8898
dc.description.abstractCre-responsive fluorescent marker alleles are powerful tools for cell lineage tracing in mice; however their utility is limited by regulation of Cre activity. When targeting hepatocytes, hydrodynamic delivery of a Cre-expression plasmid can convert Cre-responsive alleles without inducing the intracellular or systemic antiviral responses often associated with viral-derived Cre-expression vectors. In this method, rapid high-volume intravenous inoculation induces hepatocyte-targeted uptake of extracellular molecules. Here we tested whether hydrodynamic delivery of Cre protein or Cre fused to the HIV-TAT cell-penetrating peptide could convert Cre-responsive reporters in hepatocytes of mice. Hydrodynamic delivery of 2 nmol of either Cre or TAT-Cre protein converted the reporter allele in 5 to 20% of hepatocytes. Neither protein gave detectable Cre activity in endothelia, non-liver organs, or non-hepatocyte cells in liver. Using mice homozygous for a Cre-responsive marker that directs red- (Cre-naïve) or green- (Cre-converted) fluorescent proteins to the nucleus, we assessed sub-saturation Cre-activity. One month after hydrodynamic inoculation with Cre protein, 58% of hepatocyte nuclei that were green were also red, indicating that less than half of the hepatocytes that had obtained enough Cre to convert one marker allele to green were able to convert all alleles. For comparison, one month after hydrodynamic delivery of a Cre-expression plasmid with a weak promoter, only 26% of the green nuclei were also red. Our results show that hydrodynamic delivery of Cre protein allows rapid allelic conversion in hepatocytes, but Cre-activity is sub-saturating so many cells will not convert multiple Cre-responsive alleles.en_US
dc.subjectCellular biologyen_US
dc.subjectBiochemistryen_US
dc.titleHydrodynamic Delivery of Cre Proteins to Lineage-Mark or Time-Stamp Hepatocytes In Situen_US
dc.typeArticleen_US
mus.citation.extentfirstpagee91219en_US
mus.citation.issue3en_US
mus.citation.journaltitlePLoS ONEen_US
mus.citation.volume9en_US
mus.identifier.categoryEngineering & Computer Scienceen_US
mus.identifier.doi10.1371/journal.pone.0091219en_US
mus.relation.collegeCollege of Engineeringen_US
mus.relation.collegeCollege of Engineering
mus.relation.departmentMechanical & Industrial Engineering.en_US
mus.relation.universityMontana State University - Bozemanen_US


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