-245 bp of 5'-flanking region from the human platelet factor 4 gene is sufficient to drive megakaryocyte-specific expression in vivo.

Abstract:

:Platelet factor 4 (PF4) serves as a lineage-specific marker of megakaryocyte development. We previously identified two positively acting sequences in the human platelet factor 4 (hPF4) gene promoter that synergized to drive high-level luciferase reporter gene expression in vitro. Using portions of the hPF4 5'-flanking region linked to the lacZ reporter gene, we observed in this investigation that constructs with -245 bp of 5'-flanking region were more active than constructs with -2 kb of 5'-flanking region in vitro. We created two independent transgenic mouse lines with a -245-bp hPF4/lacZ construct. Cells from these mice were tested for beta-galactosidase (beta-gal) expression at the mRNA level by Northern blot and semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and at the protein level by immunohistochemistry assay. Mice from one line showed beta-gal expression specifically in all megakaryocytes of all ploidy classes from bone marrow and in platelets. Expression level was comparable to that driven by the 1.1-kb rat PF4 promoter in other transgenic mouse lines. Those in the second line showed no beta-gal expression in megakaryocytes, platelets, or any of the eight organs tested. The -245-bp hPF4 promoter is capable of driving reporter gene expression in a megakaryocyte-specific manner in transgenic mice. The small size of this megakaryocyte-specific promoter is compatible with that required in some viral vectors and may provide a model for targeting gene expression to megakaryocytes.

journal_name

Blood

journal_title

Blood

authors

Cui Z,Reilly MP,Surrey S,Schwartz E,McKenzie SE

subject

Has Abstract

pub_date

1998-04-01 00:00:00

pages

2326-33

issue

7

eissn

0006-4971

issn

1528-0020

journal_volume

91

pub_type

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