Lentiviral vectors containing mouse Csf1r control elements direct macrophage-restricted expression in multiple species of birds and mammals

Pridans, Clare; Lillico, Simon; Whitelaw, Bruce; Hume, David

doi:10.1038/mtm.2014.10

Cited by 10 publications

(20 citation statements)

References 43 publications

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“…In mammals, there is an extensive conserved STAT/IRF8 motif within FIRE, and binding of STAT1 and IRF8 has been confirmed in ChIP-seq analysis of mouse macrophages 12 . The extended STAT/IRF motifs present in mammalian FIRE are not obvious in avian FIRE (although there is a novel candidate IRF8 motif elsewhere in the element), nor are the AT-rich (FOX family) sequences in avian FIRE present in mammals, but the mouse FIRE sequence is functional as an enhancer in chicken macrophages 15 . The conservation of sequence within clades suggest that the gain and loss of motifs has been constrained within each clade on the basis that the loss of any one site produces a significant reduction in transcriptional activity, as observed in the IFNB1 enhanceosome.…”

Section: Resultsmentioning

confidence: 96%

“…The presence of FIRE is essential to the activity of a Csf1r transgenic reporter gene in mice 14 . A lentiviral vector containing FIRE and the Csf1r promoter was able to direct macrophage-restricted reporter gene expression in mouse, rat, human, pig, cow, sheep, and even chicken macrophages in vitro 15 and in transgenic sheep 16 . So, FIRE is both necessary and sufficient to direct macrophage-specific transcription from the Csf1r locus.…”

Section: Introductionmentioning

confidence: 99%

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The evolution of the macrophage-specific enhancer (Fms intronic regulatory element) within the CSF1R locus of vertebrates

Hume

Wollscheid-Lengeling

Rojo

et al. 2017

Sci Rep

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The Csf1r locus encodes the receptor for macrophage colony-stimulating factor, which controls the proliferation, differentiation and survival of macrophages. The 300 bp Fms intronic regulatory element (FIRE), within the second intron of Csf1r, is necessary and sufficient to direct macrophage-specific transcription. We have analysed the conservation and divergence of the FIRE DNA sequence in vertebrates. FIRE is present in the same location in the Csf1r locus in reptile, avian and mammalian genomes. Nearest neighbor analysis based upon this element alone largely recapitulates phylogenies inferred from much larger genomic sequence datasets. One core element, containing binding sites for AP1 family and the macrophage-specific transcription factor, PU.1, is conserved from lizards to humans. Around this element, the FIRE sequence is conserved within clades with the most conserved elements containing motifs for known myeloid-expressed transcription factors. Conversely, there is little alignment between clades outside the AP1/PU.1 element. The analysis favours a hybrid between “enhanceosome” and “smorgasbord” models of enhancer function, in which elements cooperate to bind components of the available transcription factor milieu.

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Section: Resultsmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

The evolution of the macrophage-specific enhancer (Fms intronic regulatory element) within the CSF1R locus of vertebrates

Hume

Wollscheid-Lengeling

Rojo

et al. 2017

Sci Rep

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“…Zygotes were injected with lentivirus ( Csf1r :EGFP-FIRE [12]) and developed to blastocysts in vitro to create germline transgenic Csf1r -EGFP sheep. Eight recipient ewes were transplanted with either two or three blastocysts, which resulted in three pregnancies.…”

Section: Resultsmentioning

confidence: 99%

A Csf1r-EGFP Transgene Provides a Novel Marker for Monocyte Subsets in Sheep

Pridans

Davis

Sauter

et al. 2016

The Journal of Immunology

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Expression of Csf1r in adults is restricted to cells of the macrophage lineage. Transgenic reporters based upon the Csf1r locus require inclusion of the highly conserved Fms-intronic regulatory element for expression. We have created Csf1r-EGFP transgenic sheep via lentiviral transgenesis of a construct containing elements of the mouse Fms-intronic regulatory element and Csf1r promoter. Committed bone marrow macrophage precursors and blood monocytes express EGFP in these animals. Sheep monocytes were divided into three populations, similar to classical, intermediate, and nonclassical monocytes in humans, based upon CD14 and CD16 expression. All expressed EGFP, with increased levels in the nonclassical subset. Because Csf1r expression coincides with the earliest commitment to the macrophage lineage, Csf1r-EGFP bone marrow provides a tool for studying the earliest events in myelopoiesis using the sheep as a model.

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“…CSF1R FIRE probably appeared in an early amniote, before the separation of the synapsids (mammals) and sauropsids (birds and reptiles), between 320 and 340 million years ago. We have shown elsewhere that mouse FIRE is active as a macrophage-specific enhancer in a wide range of mammals and birds ( Pridans et al, 2014 ).…”

Section: Discussionmentioning

confidence: 98%

Visualisation of chicken macrophages using transgenic reporter genes: insights into the development of the avian macrophage lineage

et al. 2014

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We have generated the first transgenic chickens in which reporter genes are expressed in a specific immune cell lineage, based upon control elements of the colony stimulating factor 1 receptor (CSF1R) locus. The Fms intronic regulatory element (FIRE) within CSF1R is shown to be highly conserved in amniotes and absolutely required for myeloid-restricted expression of fluorescent reporter genes. As in mammals, CSF1R-reporter genes were specifically expressed at high levels in cells of the macrophage lineage and at a much lower level in granulocytes. The cell lineage specificity of reporter gene expression was confirmed by demonstration of coincident expression with the endogenous CSF1R protein. In transgenic birds, expression of the reporter gene provided a defined marker for macrophage-lineage cells, identifying the earliest stages in the yolk sac, throughout embryonic development and in all adult tissues. The reporter genes permit detailed and dynamic visualisation of embryonic chicken macrophages. Chicken embryonic macrophages are not recruited to incisional wounds, but are able to recognise and phagocytose microbial antigens.

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Lentiviral vectors containing mouse Csf1r control elements direct macrophage-restricted expression in multiple species of birds and mammals

Cited by 10 publications

References 43 publications

The evolution of the macrophage-specific enhancer (Fms intronic regulatory element) within the CSF1R locus of vertebrates

The evolution of the macrophage-specific enhancer (Fms intronic regulatory element) within the CSF1R locus of vertebrates

A Csf1r-EGFP Transgene Provides a Novel Marker for Monocyte Subsets in Sheep

Visualisation of chicken macrophages using transgenic reporter genes: insights into the development of the avian macrophage lineage

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