Pf4-Cre transgenic mice allow the generation of lineage-restricted gene knockouts for studying megakaryocyte and platelet function in vivo

R Tiedt, T Schomber, H Hao-Shen, RC Skoda - Blood, 2007 - ashpublications.org
R Tiedt, T Schomber, H Hao-Shen, RC Skoda
Blood, 2007ashpublications.org
To generate transgenic mice that express Cre-recombinase exclusively in the
megakaryocytic lineage, we modified a mouse bacterial artificial chromosome (BAC) clone
by homologous recombination and replaced the first exon of the platelet factor 4 (Pf4), also
called CXCL4, with a codon-improved Cre cDNA. Several strains expressing the transgene
were obtained and one strain, Q3, was studied in detail. Crossing Q3 mice with the ROSA26-
lacZ reporter strain showed that Cre-recombinase activity was confined to megakaryocytes …
Abstract
To generate transgenic mice that express Cre-recombinase exclusively in the megakaryocytic lineage, we modified a mouse bacterial artificial chromosome (BAC) clone by homologous recombination and replaced the first exon of the platelet factor 4 (Pf4), also called CXCL4, with a codon-improved Cre cDNA. Several strains expressing the transgene were obtained and one strain, Q3, was studied in detail. Crossing Q3 mice with the ROSA26-lacZ reporter strain showed that Cre-recombinase activity was confined to megakaryocytes. These results were further verified by crossing the Q3 mice with a strain containing loxP-flanked integrin β1. Excision of this conditional allele in megakaryocytes was complete at the DNA level, and platelets were virtually devoid of the integrin β1 protein. The Pf4-Cre transgenic strain will be a valuable tool to study megakaryopoiesis, platelet formation, and platelet function.
ashpublications.org