Human <i>CYP11A1</i> promoter drives Cre recombinase expression in the brain in addition to adrenals and gonads

Wu, Hsu-Shui; Lin, Hui-Ting; Wang, Chi-Kuang Leo; Chiang, Yen-Feng; Chu, Hao-Hua; Hu, Meng Chun

doi:10.1002/dvg.20266

Cited by 16 publications

(21 citation statements)

References 39 publications

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“…Two published lines aimed at targeting steroidogenic cell lineages exhibit expression restricted to LCs within the testis (while also targeting steroidogenic organs outside of the testis such as adrenals). Use of 4.4 kb of the human Cyp11a1 promoter (which makes P 450 -SCC) has been exploited to make a SCC-iCre (Wu et al 2007). In addition to LC expression, the SCC-iCre is expressed in adrenals and the brain.…”

Section: Leydig Cellsmentioning

confidence: 99%

Good planning and serendipity: exploiting the Cre/Lox system in the testis

Smith¹

2011

Reproduction

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Over the past 20 years, genetic manipulation has revolutionised our understanding of male reproductive development and function. The advent of transgenic mouse lines has permitted elegant dissection of previously intractable issues. The development of the Cre/Lox system, which has permitted spatial and temporal localisation of genetic manipulation, has expanded upon this, and now makes up one of the primary approaches underpinning our increasing understanding of testis development and function. The success of conditional gene targeting is largely reliant upon the choice of Cre recombinase expressing mouse line, which is required to specifically target the correct cell type at the correct time. Presupposition that Cre lines will behave as expected has been one of the main oversights in the design of Cre/Lox experiments, as in practice, many Cre lines are prone to ectopic expression (both temporal and spatial), transgene silencing or genetic background effects. Empirical validation of the spatiotemporal profile of Cre expression prior to undertaking conditional gene targeting studies is essential and can be achieved through a combination of molecular and immunohistochemical approaches, along with in vivo examination of reporter gene expression in targeted tissues. This paper details the key considerations associated with exploitation of the Cre/Lox system and highlights a variety of validated Cre lines that have utility for conditional gene targeting within the testis.

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Section: Leydig Cellsmentioning

confidence: 99%

Good planning and serendipity: exploiting the Cre/Lox system in the testis

Smith¹

2011

Reproduction

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“…Recent studies using a P450scc-cre reporter (Wu, et al, 2007) have shown that the P450scc promoter is expressed in the cortex, hippocampus, thalamus, hypothalamus (dorsomedial ventromedial hypothalamus and arcuate nucleus). We prepared P450c17-GFP transgenic mice, using a 1.5 kb promoter region of the rat P450c17 gene.…”

Section: Introductionmentioning

confidence: 99%

Endogenous and synthetic neurosteroids in treatment of Niemann–Pick Type C disease

Mellon

Gong

Schonemann

2008

Brain Research Reviews

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The functions for neurosteroids during development and in response to nervous system injury are beginning to be identified. We focused on a mouse model in which we believed neurosteroid production would be altered, and which had a neurodegenerative phenotype. Niemann Pick Type-C (NP-C) is an autosomal recessive neurodegenerative disease caused by mutations in NPC1 (95%) or NPC2 (5%), resulting in lysosomal accumulation of unesterified cholesterol and glycolipids. The NIH mouse model of NP-C has a mutation in the NPC1 gene, and exhibits several pathological features of the most severe NP-C patients. How lysosomal storage and trafficking defects lead to neurodegeneration is unknown. We found that these mice had normal neurosteroidogenic enzyme activity during development, but lost this activity in the early neonatal period, prior to onset of neurological symptoms. Neurons that expressed P450scc, 3ß HSD, as well as those that expressed 3α HSD and 5α reductase were lost in adult NP-C brains, resulting in diminished concentrations of allopregnanolone. We treated NP-C mice with allopregnanolone and found that a single dose in the neonatal period resulted in a doubling of lifespan, substantial delay in onset of neurological symptoms, survival of cerebellar Purkinje and granule cell neurons, and reduction in cholesterol and ganglioside accumulation. The mechanism by which allopregnanolone elicited these effects is unknown. Our in vitro studies showed that Purkinje cell survival promoted by allopregnanolone was lost by treatment with bicuculline, suggesting GABA A receptors may play a role. We treated NP-C mice with a synthetic GABA A neurosteroid, ganaxolone (3α-hydroxy-3β-methyl-5α -pregnan-20-one). Ganaxolone treatment of NP-C mice produced beneficial neurological effects, but these effects were not as robust as those obtained using allopregnanolone. Thus, allopregnanolone may elicit its effects through GABA A receptors and through other mechanisms. Additional studies also suggest that allopregnanolone may elicit its effects through pregnane-X receptors (PXR). Our data suggest that mouse models of neurodegeneration may be beneficial in establishing both physiologic and pharmacologic actions of neurosteroids. These animal models further establish the wide range of functions of these compounds, which may ultimately be useful for treatment of human diseases.

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“…The plasmid pSCC4.4-iCreIn, which contains the 4.4-kb sequence of the human CYP11A1 promoter driving iCre gene, has been described previously (Wu et al, 2007). The transgene fragment containing 3.8 kb or 2.7 kb of CYP11A1 promoter region was generated by digestion of pSCC4.4-iCreIn with NheI and SalI, or BsrGI and SalI.…”

Section: Plasmid Constructsmentioning

confidence: 99%

“…The SCC-Cre transgenic mouse line generated previously (Wu et al, 2007) were mated with ROSA26 reporter mice to produce SCC-Cre/R26R double transgenic mice as described. To generate the transient SCC-Cre/R26R double transgenic embryos, purified transgenic DNA fragments were injected into the pronuclei of fertilized eggs derived from superovulated C57BL/6J female mated with homozygous male ROSA26 reporter mice.…”

Section: Generation Of Scc-cre/r26r Micementioning

confidence: 99%

“…Nuclear proteins Ku and Sp were further identified to bind to the À130/À94 region that is essential for the regulation of Cyp11a1 promoter activity in C6 cells (Zhang et al, 1995;Hammer et al, 2004). We recently generated SCC-Cre transgenic mice in which Cre recombinase expression is under the regulation of the 4.4-kb promoter from human CYP11A1 gene (Wu et al, 2007). In SCC-Cre mice, Cre expression was found in specific regions of the brain through a Cre-loxP-based method (Soriano, 1999;Kwan, 2002), which indicated that the 4.4-kb 5 0 -flanking sequence contains essential regulatory elements for CYP11A1 expression in the brain.…”

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confidence: 97%

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Differential regulation of the human CYP11A1 promoter in mouse brain and adrenals

Chiang

Lin

et al. 2011

Journal Cellular Physiology

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CYP11A1 encodes the first enzyme of steroid biosynthesis, cytochrome P450scc. The expression of CYP11A1 in the nervous system allows neurosteroids to be synthesized de novo. In the classic steroidogenic tissues, adrenals and gonads, the key regulator controlling CYP11A1 expression is steroidogenic factor-1 (SF-1), but the transcriptional regulation of CYP11A1 in the brain is unclear. We recently used the 4.4-kb regulatory region of the human CYP11A1 gene to drive Cre recombinase expression in the diencephalon and midbrain. In this study, we characterized the regional-specific expression of Cre reporter in the SCC-Cre transgenic brain using a transient Cre/ROSA26R transgenic system. Mutation of either the upstream or proximal SF-1 binding site did not affect brain CYP11A1 promoter activity. The upstream SF-1 binding site, however, is required for CYP11A1 promoter function in the embryonic adrenals. The 3.8-kb promoter, like the 4.4-kb length promoter, directed Cre expression in the diencephalon, midbrain and olfactory epithelium, whereas Cre expression controlled by the 2.7-kb promoter was only observed in the caudal part of midbrain. This suggests that the 5'-flanking region between 3.8 and 2.7 kb contains a crucial element for activation of CYP11A1 promoter in the diencephalon, olfactory epithelium and the anterior part of midbrain. Thus we have identified regions of the promoter that control CYP11A1 expression in the brain and embryonic adrenals.

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Human CYP11A1 promoter drives Cre recombinase expression in the brain in addition to adrenals and gonads

Cited by 16 publications

References 39 publications

Good planning and serendipity: exploiting the Cre/Lox system in the testis

Good planning and serendipity: exploiting the Cre/Lox system in the testis

Endogenous and synthetic neurosteroids in treatment of Niemann–Pick Type C disease

Differential regulation of the human CYP11A1 promoter in mouse brain and adrenals

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