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A genetic strategy for single and combinatorial analysis of miRNA function in mammalian hematopoietic stem cells.

Papapetrou, Eirini P., Korkola, James E., Sadelain, Michel·2010·Stem Cells
DOI 10.1002/stem.257
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Abstract

The regulatory role of micro-RNAs (miRNAs) in hematopoietic development is increasingly appreciated. Reverse genetics strategies based on the targeted disruption of miRNAs offer a powerful tool to study miRNA functions in mammalian hematopoiesis. The miR-144/451 cluster comprises two miRNAs coexpressed from a common precursor transcript in an erythroid-specific manner. To decipher the contribution of each miRNA of the cluster in mammalian erythropoiesis, we developed a strategy for stable in vivo individual and combinatorial miRNA inhibition. We developed decoy target sequences for each miRNA expressed by lentiviral vectors marked with distinct fluorescent proteins and used them to probe the functions of miR-144 and miR-451 in the murine hematopoietic system in a competitive repopulation setting. Murine hematopoietic chimeras expressing lentiviral-encoded inhibitory sequences specific for miR-144 or miR-451 exhibited markedly reduced Ter119+ erythroblast counts, with the combined knockdown showing additive effect. These chimeras showed abnormal patterns of erythroid differentiation primarily affecting the proerythroblast to basophilic erythroblast transition, coinciding with the stage where expression of the miRNA cluster is dramatically induced and posttranscriptional gene regulation becomes prominent. These results reveal a role for the miR-144/451 locus in mammalian erythropoiesis and provide the first evidence of functional cooperativity between clustered miRNAs in the hematopoietic system. The strategy described herein will prove useful in functional miRNA studies in mammalian hematopoietic stem cells.

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Crossref · 10.1002/stem.257

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  • miR-451 and miR-144 are crucial to erythropoiesis.

    “Extensive evidence has shown that miR-451 and miR-144 are crucial to erythropoiesis and are located within the same gene cluster and regulated by the erythrocyte-specific transcription factor GATA1”

    From Red Blood Cells as Potential Repositories of MicroRNAs in the Circulatory System (2020) · cites this paper as Papapetrou et al., 2010

  • miR-200a and miR-150 are involved in erythrocyte differentiation.

    “multiple miRNAs such as miR-200a and miR-150 have also been shown to be involved in the process of erythrocyte differentiation”

    From Red Blood Cells as Potential Repositories of MicroRNAs in the Circulatory System (2020) · cites this paper as Papapetrou et al., 2010

  • miR-451 and miR-144 are located within the same gene cluster and regulated by GATA1.

    “Extensive evidence has shown that miR-451 and miR-144 are crucial to erythropoiesis and are located within the same gene cluster and regulated by the erythrocyte-specific transcription factor GATA1”

    From Red Blood Cells as Potential Repositories of MicroRNAs in the Circulatory System (2020) · cites this paper as Papapetrou et al., 2010

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Metadata

Authors
Papapetrou, E. P., Korkola, J. E., & Sadelain, M.
Affiliations
  • Center for Cell Engineering, Memorial Sloan-Kettering Cancer Center, New York, New York, USA
  • Cell Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, USA
Venue
Stem Cells
Publisher
Oxford University Press (OUP)
Published
2009-11-12
Volume
28
Issue
2
Pages
287-296
ISSN
  • 1066-5099
  • 1549-4918
DOI
10.1002/stem.257
Cited by
73
References
3
Language
en
Licence
academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
Funders
  • NIH (HL 57612, PO1 CA059350)
  • New York State Stem Cell Science (NYSTEM-016)

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