Distinct regulatory elements direct Delta1 expression in the nervous system and paraxial mesoderm of transgenic mice

The expression of prosaposin is temporally and spatially regulated at transcriptional and post-translational levels. Transgenic mice with various 5′-flanking deletions of the prosaposin promoter fused to luciferase (LUC) reporters were used to define its temporal regulatory region. LUC expression in the transgenic mice carrying constructs with 234bp (234LUC), 310bp (310LUC) or 2400bp (2400LUC) of the 5′-flanking region was analysed in the central nervous system and eye throughout development. For 310LUC and 2400LUC, low-level LUC activity was maintained until embryonal day 18 in brain, eye and spinal cord. The peak level of LUC activity was at birth, with return to a plateau (1/3 of peak) throughout adulthood. Deletion of the region that included the retinoic acid-receptor-related orphan receptor (RORα)-binding site and sequence-specific transcription factor (Sp1) cluster sites (44—310bp) suppressed the peak of activity. By comparison, the peak level for 234LUC was shifted 2 weeks into neonatal life in the brain, but not in the eye, and no peak of activity was observed in the spinal cord. The endogenous prosaposin mRNA in eye, spinal cord and cerebellum had low-level expression before birth and continued to increase into adulthood. In cerebrum, the endogenous mRNA showed similar expression profile to constructs 310LUC, 2400LUC and 234LUC, with the peak expression at 1 week and a decreased level in adult. In the brain of the newborn, 2400LUC was highly expressed in the trigeminal ganglion and brain stem regions when compared with the generalized expression pattern for endogenous prosaposin mRNA. These results suggest that the modifiers (RORα- and Sp1-binding sites) residing within 310bp of the 5′-flanking region mediate developmental regulation in the central nervous system and eye. Additional regulatory elements outside the 5′ region of the 2400bp promoter fragment appear to be essential for the physiological control of the prosaposin locus.

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Autoregulation and multiple enhancers control Math1 expression in the developing nervous system

Development ◽

10.1242/dev.127.6.1185 ◽

2000 ◽

Vol 127 (6) ◽

pp. 1185-1196 ◽

Cited By ~ 19

Author(s):

A.W. Helms ◽

A.L. Abney ◽

N. Ben-Arie ◽

H.Y. Zoghbi ◽

J.E. Johnson

Keyword(s):

Nervous System ◽

Transgenic Mice ◽

Neural Tube ◽

Hair Cells ◽

Neuronal Cell ◽

Cell Types ◽

Regulatory Elements ◽

Coding Region ◽

Consensus Binding Site ◽

Auditory Systems

Development of the vertebrate nervous system requires the actions of transcription factors that establish regional domains of gene expression, which results in the generation of diverse neuronal cell types. MATH1, a transcription factor of the bHLH class, is expressed during development of the nervous system in multiple neuronal domains, including the dorsal neural tube, the EGL of the cerebellum and the hair cells of the vestibular and auditory systems. MATH1 is essential for proper development of the granular layer of the cerebellum and the hair cells of the cochlear and vestibular systems, as shown in mice carrying a targeted disruption of Math1. Previously, we showed that 21 kb of sequence flanking the Math1-coding region is sufficient for Math1 expression in transgenic mice. Here we identify two discrete sequences within the 21 kb region that are conserved between mouse and human, and are sufficient for driving a lacZ reporter gene in these domains of Math1 expression in transgenic mice. The two identified enhancers, while dissimilar in sequence, appear to have redundant activities in the different Math1 expression domains except the spinal neural tube. The regulatory mechanisms for each of the diverse Math1 expression domains are tightly linked, as separable regulatory elements for any given domain of Math1 expression were not found, suggesting that a common regulatory mechanism controls these apparently unrelated domains of expression. In addition, we demonstrate a role for autoregulation in controlling the activity of the Math1 enhancer, through an essential E-box consensus binding site.

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The CD11b promoter directs high-level expression of reporter genes in macrophages in transgenic mice [published erratum appears in Blood 1995 Apr 1;85(7):1983]

Blood ◽

10.1182/blood.v85.2.319.319 ◽

1995 ◽

Vol 85 (2) ◽

pp. 319-329 ◽

Cited By ~ 66

Author(s):

S Dziennis ◽

RA Van Etten ◽

HL Pahl ◽

DL Morris ◽

TL Rothstein ◽

...

Keyword(s):

Gene Expression ◽

Transgenic Mice ◽

Transgene Expression ◽

Heterologous Gene Expression ◽

Myeloid Cells ◽

Reporter Genes ◽

Regulatory Elements ◽

Specific Expression ◽

High Level

Abstract CD11b is the alpha chain of the Mac-1 integrin and is preferentially expressed in myeloid cells (neutrophils, monocytes, and macrophages). We have previously shown that the CD11b promoter directs cell-type- specific expression in myeloid lines using transient transfection assays. To confirm that these promoter sequences contain the proper regulatory elements for correct myeloid expression of CD11b in vivo, we have used the -1.7-kb human CD11b promoter to direct reporter gene expression in transgenic mice. Stable founder lines were generated with two different reporter genes, a Thy 1.1 surface marker and the Escherichia coli lacZ (beta-galactosidase) gene. Analysis of founders generated with each reporter demonstrated that the CD11b promoter was capable of driving high levels of transgene expression in murine macrophages for the lifetime of the animals. Similar to the endogenous gene, transgene expression was preferentially found in mature monocytes, macrophages, and neutrophils and not in myeloid precursors. These experiments indicate that the -1.7 CD11b promoter contains the regulatory elements sufficient for high-level macrophage expression. This promoter should be useful for targeting heterologous gene expression to mature myeloid cells.

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Distant regulatory elements in a Sox10-βGEO BAC transgene are required for expression ofSox10in the enteric nervous system and other neural crest-derived tissues

Developmental Dynamics ◽

10.1002/dvdy.20769 ◽

2006 ◽

Vol 235 (5) ◽

pp. 1413-1432 ◽

Cited By ~ 48

Author(s):

Karen K. Deal ◽

V. Ashley Cantrell ◽

Ronald L. Chandler ◽

Thomas L. Saunders ◽

Douglas P. Mortlock ◽

...

Keyword(s):

Nervous System ◽

Neural Crest ◽

Enteric Nervous System ◽

Regulatory Elements ◽

Bac Transgene

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Transgenic mice expressing the Peripherin-EGFP genomic reporter display intrinsic peripheral nervous system fluorescence

Transgenic Research ◽

10.1007/s11248-008-9210-7 ◽

2008 ◽

Vol 17 (6) ◽

pp. 1103-1116 ◽

Cited By ~ 8

Author(s):

Samuel McLenachan ◽

Yona Goldshmit ◽

Kerry J. Fowler ◽

Lucille Voullaire ◽

Timothy P. Holloway ◽

...

Keyword(s):

Nervous System ◽

Transgenic Mice ◽

Peripheral Nervous System

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Fast skeletal muscle-specific expression of a quail troponin I gene in transgenic mice.

Molecular and Cellular Biology ◽

10.1128/mcb.8.12.5072 ◽

1988 ◽

Vol 8 (12) ◽

pp. 5072-5079 ◽

Cited By ~ 7

Author(s):

P L Hallauer ◽

K E Hastings ◽

A C Peterson

Keyword(s):

Skeletal Muscle ◽

Transgenic Mice ◽

Troponin I ◽

Fiber Type ◽

Regulatory Elements ◽

Evolutionary Divergence ◽

Mrna Levels ◽

Fast Skeletal Muscle ◽

Specific Expression ◽

Exon 2

We have produced seven lines of transgenic mice carrying the quail gene encoding the fast skeletal muscle-specific isoform of troponin I (TnIf). The quail DNA included the entire TnIf gene, 530 base pairs of 5'-flanking DNA, and 1.5 kilobase pairs of 3'-flanking DNA. In all seven transgenic lines, normally initiated and processed quail TnIf mRNA was expressed in skeletal muscle, where it accumulated to levels comparable to that in quail muscle. Moreover, in the three lines tested, quail TnIf mRNA levels were manyfold higher in a fast skeletal muscle (gastrocnemius) than in a slow skeletal muscle (soleus). We conclude that the cellular mechanisms directing muscle fiber type-specific TnIf gene expression are mediated by cis-regulatory elements present on the introduced quail DNA fragment and that they control TnIf expression by affecting the accumulation of TnIf mRNA. These elements have been functionally conserved since the evolutionary divergence of birds and mammals, despite the major physiological and morphological differences existing between avian (tonic) and mammalian (twitch) slow muscles. In lines of transgenic mice carrying multiple tandemly repeated copies of the transgene, an aberrant quail TnIf transcript (differing from normal TnIf mRNA upstream of exon 2) also accumulated in certain tissues, particularly lung, brain, spleen, and heart tissues. However, this aberrant transcript was not detected in a transgenic line which carries only a single copy of the quail gene.

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Fluorescent protein expression driven by her4 regulatory elements reveals the spatiotemporal pattern of Notch signaling in the nervous system of zebrafish embryos

Developmental Biology ◽

10.1016/j.ydbio.2006.10.020 ◽

2007 ◽

Vol 301 (2) ◽

pp. 555-567 ◽

Cited By ~ 92

Author(s):

Sang-Yeob Yeo ◽

MinJung Kim ◽

Hyung-Seok Kim ◽

Tae-Lin Huh ◽

Ajay B. Chitnis

Keyword(s):

Nervous System ◽

Protein Expression ◽

Notch Signaling ◽

Fluorescent Protein ◽

Regulatory Elements ◽

Spatiotemporal Pattern ◽

Zebrafish Embryos

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Distal regulatory elements from the mouse metallothionein locus stimulate gene expression in transgenic mice

Molecular and Cellular Biology ◽

10.1128/mcb.13.9.5266-5275.1993 ◽

1993 ◽

Vol 13 (9) ◽

pp. 5266-5275

Author(s):

R D Palmiter ◽

E P Sandgren ◽

D M Koeller ◽

R L Brinster

Keyword(s):

Transgenic Mice ◽

Regulatory Elements ◽

High Level Expression ◽

Hypersensitive Sites ◽

Enhanced Expression ◽

Rat Growth ◽

Flanking Sequences ◽

High Level ◽

Distal Regulatory Elements ◽

Flanking Regions

DNA regions of 10 and 7 kb that flank the mouse metallothionein II (MT-II) and MT-I genes, respectively, were combined with a minimally marked MT-I (MT-I*) gene and tested in transgenic mice. This construct resulted in (i) position-independent expression of MT-I* mRNA and copy number-dependent expression, (ii) levels of hepatic MT-I mRNA per cell per transgene that were about half that derived from endogenous MT-I genes, (iii) appropriate regulation by metals and hormones, and (iv) tissue distribution of transgene mRNA that resembled that of endogenous MT-I mRNA. These features were not observed when MT-I* was tested without the flanking regions. These MT-I flanking sequences also improved the expression of rat growth hormone reporter genes, with or without introns, that were under the control of the MT-I promoter. Moreover, they enhanced expression from two of four heterologous promoters/enhancers that were tested. Deletion analysis indicated that regions known to have DNase I-hypersensitive sites were necessary but not sufficient for high-level expression. These data suggest that the DNA regions flanking the mouse MT-I and MT-II genes have functions like the locus control regions described for other genes.

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Patterning the ascidian nervous system: structure, expression and transgenic analysis of the CiHox3 gene

Development ◽

10.1242/dev.126.21.4737 ◽

1999 ◽

Vol 126 (21) ◽

pp. 4737-4748 ◽

Cited By ~ 1

Author(s):

A. Locascio ◽

F. Aniello ◽

A. Amoroso ◽

M. Manzanares ◽

R. Krumlauf ◽

...

Keyword(s):

Central Nervous System ◽

Nervous System ◽

Hox Genes ◽

Regional Identity ◽

Regulatory Elements ◽

System Structure ◽

Anteroposterior Patterning ◽

Hox Gene ◽

The Central Nervous System ◽

Group 3

Hox genes play a fundamental role in the establishment of chordate body plan, especially in the anteroposterior patterning of the nervous system. Particularly interesting are the anterior groups of Hox genes (Hox1-Hox4) since their expression is coupled to the control of regional identity in the anterior regions of the nervous system, where the highest structural diversity is observed. Ascidians, among chordates, are considered a good model to investigate evolution of Hox gene, organisation, regulation and function. We report here the cloning and the expression pattern of CiHox3, a Ciona intestinalis anterior Hox gene homologous to the paralogy group 3 genes. In situ hybridization at the larva stage revealed that CiHox3 expression was restricted to the visceral ganglion of the central nervous system. The presence of a sharp posterior boundary and the absence of transcript in mesodermal tissues are distinctive features of CiHox3 expression when compared to the paralogy group 3 in other chordates. We have investigated the regulatory elements underlying CiHox3 neural-specific expression and, using transgenic analysis, we were able to isolate an 80 bp enhancer responsible of CiHox3 activation in the central nervous system (CNS). A comparative study between mouse and Ciona Hox3 promoters demonstrated that divergent mechanisms are involved in the regulation of these genes in vertebrates and ascidians.

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A 2-kb c-mpl promoter fragment is sufficient to direct expression to the megakaryocytic lineage and sites of embryonic hematopoiesis in transgenic mice

Blood ◽

10.1182/blood-2002-01-0281 ◽

2002 ◽

Vol 100 (3) ◽

pp. 1072-1074 ◽

Cited By ~ 13

Author(s):

Sandra Ziegler ◽

Kurt Bürki ◽

Radek C. Skoda

Keyword(s):

Transgenic Mice ◽

Reporter Gene ◽

Fetal Liver ◽

Regulatory Elements ◽

Yolk Sac ◽

Placental Alkaline Phosphatase ◽

Hematopoietic Progenitors ◽

Thrombopoietin Receptor ◽

Human Placental Alkaline Phosphatase ◽

Dna Fragment

Abstract Thrombopoietin receptor c-mpl is expressed on hematopoietic progenitors and cells of the megakaryocytic lineage. The c-mpl promoter may, therefore, be useful for directing the expression of transgenes. We tested whether a 2-kb genomic DNA fragment comprising the putative c-mpl regulatory elements and most of the 5′-untranslated region of mouse c-mpl is able to direct the expression of a reporter gene to hematopoietic cells in transgenic mice. As a reporter gene we used the human placental alkaline phosphatase (PLAP). In adult transgenic mice, PLAP expression was specifically detected in megakaryocytes and platelets. Embryos showed PLAP reporter gene expression already in the yolk sac at embryonic day 6.5 (E6.5) and in blood islands at E7.5. At E9.5, expression was found in blood vessels of the yolk sac and the embryo proper, followed by high levels of expression in the fetal liver at E11.5. Expression in E6.5 yolk sac is compatible with a function of c-mpl and its ligand, thrombopoietin, in the earliest stages of embryonic hematopoiesis.

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