Identification of hematopoietic-specific regulatory elements from the CD45 gene and use for lentiviral tracking of transplanted cells

Khanh L. Duong, Satyabrata Das, Shuyang Yu, Jennifer Y. Barr, Snehalata Jena, Eunmi Kim, Nicolas Zavazava, John D. Colgan, Hai Hui Xue, Dana N. Levasseur

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


The development of a hematopoietic reporter is crucial for determining the fate of lineages derived from cell-based therapies. A marking system will enable safer embryonic stem and induced pluripotent stem cell-based derivation of blood lineages and facilitate the development of efficient cellular reprogramming strategies based on direct fibroblast conversion. Here we report that the protein tyrosine phosphatase CD45 is an ideal candidate gene on which to base a hematopoietic reporter. CD45 regulatory elements were discovered by analyzing transcription factor chromatin occupancy (ChIP-seq) and promoter nuclease sensitivity (DNase-seq) to identify minimally sufficient sequences required for expression. After cloning the CD45 regulatory elements into an attenuated lentiviral backbone, we found that two transcriptional initiation regions were essential for high-level expression. Expressing CD45 promoters containing these regions and tethered to green fluorescent protein (GFP) in a primary B-cell differentiation assay and a transplantation model resulted in high levels of GFP in lymphoid, myeloid, and nucleated erythroid cells in mouse and human blood cell lineages. Moreover, GFP levels remained high 5months after secondary transplantation, indicating persistence of the reporter. No CD45-driven GFP expression is observed after fibroblast or embryonic stem cell transduction. The GFP reporter is seen only after embryonic stem cells differentiateinto hematopoietic cell progenitors and lineages, suggesting that this hematopoietic reportersystem could be useful in validating potential autologous blood cell therapies.

Original languageEnglish (US)
Pages (from-to)761-772.e10
JournalExperimental Hematology
Issue number9
StatePublished - 2014

Bibliographical note

Funding Information:
This work was supported by a Basil O'Connor Starter Scholar Research Award Grant (5-FY10-45) from the March of Dimes, startup funds from the Carver College of Medicine Department of Internal Medicine, and the University of Iowa Graduate College .

Publisher Copyright:
© 2014 ISEH - International Society for Experimental Hematology.


Dive into the research topics of 'Identification of hematopoietic-specific regulatory elements from the CD45 gene and use for lentiviral tracking of transplanted cells'. Together they form a unique fingerprint.

Cite this