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Introduction
Sandra Piltz currently works at the South Australian Health and Medical Research Institute (SAHMRI) as part of the University of Adelaide. Sandra does research in Genetics, Molecular Biology and Cell Biology. Her most recent publication is 'Knockout of the epilepsy gene Depdc5 in mice causes severe embryonic dysmorphology with hyperactivity of mTORC1 signalling.'
Skills and Expertise
Additional affiliations
April 2008 - present
Publications
Publications (83)
Background
The development of sequence-specific precision treatments like CRISPR gene editing therapies for Duchenne muscular dystrophy (DMD) requires sequence humanized animal models to enable the direct clinical translation of tested strategies. The current available integrated transgenic mouse model containing the full-length human DMD gene, Tg(...
CRISPR-Cas9 technology has facilitated development of strategies that can potentially provide more humane and effective methods to control invasive vertebrate species, such as mice. One promising strategy is X chromosome shredding which aims to bias offspring towards males, resulting in a gradual and unsustainable decline of females. This method ha...
The development of sequence-specific precision treatments like CRISPR gene-editing therapies for Duchenne Muscular Dystrophy (DMD) requires sequence humanised animal models to enable the direct clinical translation of tested strategies. The current available integrated transgenic mouse model containing the full-length human DMD gene, Tg(DMD)72Thoen...
Mutations in the X-linked gene PCDH19 are the cause of PCDH19-Clustering epilepsy, an infantile-onset disorder characterized by seizures and intellectual disabilities. Although several intra and extracellular functions of PCDH19 have been identified, the spatiotemporal impact of Pcdh19 deletion in vivo is poorly understood. To investigate the conse...
We implicated the X-chromosome THOC2 gene, which encodes the largest subunit of the highly-conserved TREX (Transcription-Export) complex, in a clinically complex neurodevelopmental disorder with intellectual disability as the core phenotype. To study the molecular pathology of this essential eukaryotic gene, we generated a mouse model based on a hy...
CRISPR-Cas9 technology has facilitated development of strategies that can potentially provide more humane and effective methods to control invasive vertebrate species, such as mice. One promising strategy is X chromosome shredding which aims to bias offspring towards males, resulting in a gradual and unsustainable decline of females. This method ha...
Transcriptional enhancer elements are responsible for orchestrating the temporal and spatial control over gene expression that is crucial for programming cell identity during development1–3. Here we describe a novel enhancer element that is important for regulating the expression of Prox1 in lymphatic endothelial cells. This evolutionarily conserve...
Invasive rodents are a major cause of environmental damage and biodiversity loss, particularly on islands. Unlike insects, genetic biocontrol strategies including population-suppressing gene drives with biased inheritance have not been developed in mice. Here, we demonstrate a gene drive strategy (tCRISPR) that leverages super-Mendelian transmissio...
Developmental and epileptic encephalopathies (DEE) are characterized by pharmacoresistant seizures with concomitant intellectual disability. Epilepsy of infancy with migrating focal seizures (EIMFS) is one of the most severe of these syndromes. De novo variants in ion channels, including gain-of-function variants in KCNT1, have been found to play a...
Gene drives are genetic elements that are transmitted to greater than 50% of offspring and have potential for population modification or suppression. While gene drives are known to occur naturally, the recent emergence of CRISPR-Cas9 genome-editing technology has enabled generation of synthetic gene drives in a range of organisms including mosquito...
Invasive rodents, including house mice, are a major cause of environmental damage and biodiversity loss, particularly in island ecosystems. Eradication can be achieved through the distribution of rodenticide, but this approach is expensive to apply at scale, can have negative impacts (e.g. on non-target species, or through contamination), has anima...
Central conducting lymphatic anomaly (CCLA), characterized by the dysfunction of core collecting lymphatic vessels including the thoracic duct and cisterna chyli, and presenting as chylothorax, pleural effusions, chylous ascites, and lymphedema, is a severe disorder often resulting in fetal or perinatal demise. Although pathogenic variants in RAS/m...
Enhancers are vitally important during embryonic development to control the spatial and temporal expression of genes. Recently, large scale genome projects have identified a vast number of putative developmental regulatory elements. However, the proportion of these that have been functionally assessed is relatively low. While enhancers have traditi...
Precise genomic modification using prime editing (PE) holds enormous potential for research and clinical applications. In this study, we generated all-in-one prime editing (PEA1) constructs that carry all the components required for PE, along with a selection marker. We tested these constructs (with selection) in HEK293T, K562, HeLa and mouse embry...
Precise genomic modification using prime editing (PE) holds enormous potential for research and clinical applications. Currently, the delivery of PE components to mammalian cell lines requires multiple plasmid vectors. To overcome this limitation, we generated all-in-one prime editing (PEA1) constructs that carry all the components required for PE,...
Enhancers are vitally important during embryonic development to control the spatial and temporal expression of genes. Recently, large scale genome projects have identified a vast number of putative developmental regulatory elements. However, the proportion of these that have been functionally assessed is relatively low. While enhancers have traditi...
Developmental and epileptic encephalopathies (DEE) are characterized by pharmacoresistant seizures with concomitant intellectual disability. Epilepsy of infancy with migrating focal seizures (EIMFS) is one of the most severe of these syndromes. De novo mutations in ion channels, including gain-of-function variants in KCNT1 , have been found to play...
Animal models are needed to develop interventions to prevent or treat intrauterine growth restriction (IUGR). Fetal growth rates and effects of in utero exposures differ between sexes, but little is known about sex-specific effects of increasing litter size. We established a murine IUGR model using pregnancies generated by multiple embryo transfers...
CRISPR-based synthetic gene drives have the potential to deliver a more effective and humane method of invasive vertebrate pest control than current strategies. Relatively efficient CRISPR gene drive systems have been developed in insects and yeast but not in mammals. Here, we investigated the efficiency of CRISPR-Cas9-based gene drives in Mus musc...
CRISPR-based synthetic gene drives have the potential to deliver a more effective and humane method of invasive vertebrate pest control than current strategies. Relatively efficient CRISPR gene drives have been developed in insects and yeast but not in mammals. Here we investigated the efficiency of CRISPR/Cas9-based gene drives in Mus musculus by...
Sotos syndrome is a developmental disorder characterized by a suite of clinical features. In children, the three cardinal features of Sotos syndrome are a characteristic facial appearance, learning disability, and overgrowth (height and/or head circumference > 2 standard deviations above average). These features are also evident in adults with this...
Background:
CRISPR-Cas9 gene-editing technology has facilitated the generation of knockout mice, providing an alternative to cumbersome and time-consuming traditional embryonic stem cell-based methods. An earlier study reported up to 16% efficiency in generating conditional knockout (cKO or floxed) alleles by microinjection of 2 single guide RNAs...
The RNA-guided endonuclease CRISPR-Cas system from Streptococcus pyogenes (SpCas9) is widely used for generating genetically modified mice via zygotic microinjection. Although SpCas9 is a potent mutagen, it requires an NGG proto-spacer adjacent motif (PAM) at the target site, restricting sequence targetability. Here, we show that RNA-guided endonuc...
CRISPR–Cas9 gene editing technology has considerably facilitated the generation of mouse knockout alleles, relieving many of the cumbersome and time-consuming steps of traditional mouse embryonic stem cell technology. However, the generation of conditional knockout alleles remains an important challenge. An earlier study reported up to 16% efficien...
PCDH19-Girls Clustering Epilepsy (PCDH19-GCE) is a childhood epileptic encephalopathy characterised by a spectrum of neurodevelopmental problems. PCDH19-GCE is caused by heterozygous loss-of-function mutations in the X-chromosome gene, Protocadherin 19 (PCDH19) encoding a cell-cell adhesion molecule. Intriguingly, hemizygous males are generally una...
X-linked diseases typically exhibit more severe phenotypes in males than females. In contrast, protocadherin 19 (PCDH19) mutations cause epilepsy in heterozygous females but spare hemizygous males. The cellular mechanism responsible for this unique pattern of X-linked inheritance is unknown. We show that PCDH19 contributes to adhesion specificity i...
DEPDC5 mutations have recently been shown to cause epilepsy in humans. Evidence from in vitro studies has implicated DEPDC5 as a negative regulator of mTORC1 during amino acid insufficiency as part of the GATOR1 complex. To investigate the role of DEPDC5 in vivo we generated a null mouse model using targeted CRISPR mutagenesis. Depdc5 homozygotes d...
X-linked diseases typically exhibit more severe phenotypes in males than females. In contrast, Protocadherin 19 ( PCDH19 ) mutations cause epilepsy in heterozygous females but spare hemizygous males. The cellular mechanism responsible for this unique pattern of X-linked inheritance is unknown. We show that PCDH19 contributes to highly specific comb...
Protocadherin 19 (Pcdh19) is an X-linked gene belonging to the protocadherin superfamily, whose members are predominantly expressed in the central nervous system and have been implicated in cell-cell adhesion, axon guidance and dendrite self-avoidance. Heterozygous loss-of-function mutations in humans result in the childhood epilepsy disorder PCDH1...
Genetically modified mice are an invaluable resource for investigating gene function and identifying the molecular pathology of disease. However, generating mutant mice by modification of Embryonic Stem cells and subsequent breeding of chimeric mice is time consuming and labour intensive. Recent advances in genome editing technology using the Clust...
Genetically modified mice are an invaluable resource for investigating gene function and identifying the molecular pathology of disease. However, generating mutant mice by modification of Embryonic Stem cells and subsequent breeding of chimeric mice is time consuming and labour intensive. Recent advances in genome editing technology using the Clust...
Although CRISPR-CAS9 genome editing technology is still in its infancy, it has already begun to transform biomedical research. One area of enormous potential is rapid and cost-effective generation of genetic mouse models. Over the past 12 months we have assessed the efficacy of in vivo genome editing by injection of C57BL/6 mouse zygotes with CRISP...
SOX3-26ala protein is cleared from neuroprogenitor cells in vivo. Sox3-26ala<->WT chimeras were cut at 7.5 dpc (transverse section through neural groove), 9.5 dpc (transverse section through neural tube) or 10.5 dpc (sagittal section through ventral diencephalon) and stained with antibodies against SOX3 and NEO. Mutant Sox3-26ala cells (NEO positiv...
SOX3-26ala expressing cells show no signs of apoptosis. Coronal section of a 13.5 dpc Sox3-26ala<->WT chimera stained with antibodies against SOX3 and Activated Caspase3 (BD Pharmingen, #559565, 1∶1000). Boxed area in overlay shows a rare Activated Caspase3 positive cell, while the arrowhead indicates a patch of mutant cells (low SOX3) in which no...
A) Phospor image of an in vitro transcription/translation reaction of mouse and human SOX3 WT and 26ala expression constructs. SOX3 protein of the expected size is indicated by the arrow. Two strong bands were generated by the mouse plasmids, possibly reflecting the inclusion of an upstream ATG in the plasmid, while human plasmids generated only on...
Mutant SOX3-26ala protein aggregates in vitro. COS cells were transfected with mouse or human pcDNA3.1 SOX3 or pcDNA3.1 SOX3-26ala expression plasmids and two days later cells were fixed and stained with a SOX3 antibody (Green) and DAPI (Blue). Cells were scored according to the localisation of the SOX3 staining as nuclear, cytoplasmic or peri-nucl...
Polyalanine expansions in transcription factors have been associated with eight distinct congenital human diseases. It is thought that in each case the polyalanine expansion causes misfolding of the protein that abrogates protein function. Misfolded proteins form aggregates when expressed in vitro; however, it is less clear whether aggregation is o...
Congenital hydrocephalus (CH) is a life-threatening medical condition in which excessive accumulation of CSF leads to ventricular expansion and increased intracranial pressure. Stenosis (blockage) of the Sylvian aqueduct (Aq; the narrow passageway that connects the third and fourth ventricles) is a common form of CH in humans, although the genetic...
SOX3 expression, apoptosis analysis and ChP morphology in wild type and Sox3 transgenic embryos. A–H: SOX3 immunohistochemical analysis of wild type (A,B,E,F) and Nr/+ (C,D,G,H) embryos. In wild type embryos, a lower level of SOX3 was evident in the SCO at 15.5 dpc (A,B) and 18.5 dpc (E,F) in comparison to flanking periluminal cells. This differenc...
Characterisation of CH phenotype in Sox3 transgenic mice. A,B: H&E stained sagittal sections of P26 brains showing that the fourth ventricle was not expanded in Nr/+ CH mice (B) in comparison to wild type (A). C–F: Immunohistochemical analysis of SOX3 (red; C,E) and EGFP (green; D,F) expression in wild type (C,D) and Nr/+ (E,F)18.5 dpc coronally se...
We are studying families with an inherited form of pituitary
growth hormone deficiency termed X-linked Hypopituitarism
(XH) in which only boys are affected. In addition to poor
pituitary function, some boys with XH also have mild
intellectual disability. We have previously shown that XH is
due to an unusual change in the SOX3 gene in which the
numb...
We are studying families with an inherited form of pituitary growth hormone deficiency termed X-linked Hypopituitarism (XH) in which only boys are affected. In addition to poor pituitary function, some boys with XH also have mild intellectual disability. We have previously shown that XH is due to an unusual change in the SOX3 gene in which the numb...
List of 413,494 unique tags.
List of unique tags mapped to RefSeq sequences based on 3' end sequences.
List of unique tags mapped to redundant mouse ESTs based on 3' end sequences.
List of unique tags mapped to miRNAs, pre-miRNAs or miRNA-star based on 3' end sequences.
List of unique tags mapped to RefSeq sequences based on 5' end sequences.
List of unique tags mapped to redundant mouse ESTs based on 5' end sequences.
List of unique tags mapped to a single locus in the mouse genome based on 5' end sequences.
List of unique tags mapped to multiple loci in the mouse genome based on 3' end sequences.
List of unique tags mapped to multiple loci in the mouse genome based on 5' end sequences.
List of unique tags mapped to repetitive elements and ncRNAs based on the 3' end sequences.
List of unique tags mapped to repetitive elements and ncRNAs based on the 5' end sequences.
List of unique tags mapped to a single locus in the mouse genome based on 3' end sequences.
List of known miRNAs in the E15.5 developing mouse brain.
Supplementary figures and data for statistical analysis.
MicroRNAs (miRNAs) are small non-coding RNAs that can exert multilevel inhibition/repression at a post-transcriptional or protein synthesis level during disease or development. Characterisation of miRNAs in adult mammalian brains by deep sequencing has been reported previously. However, to date, no small RNA profiling of the developing brain has be...
The Lmo2 gene encodes a transcriptional cofactor critical for the development of hematopoietic stem cells. Ectopic LMO2 expression causes leukemia in T-cell acute lymphoblastic leukemia (T-ALL) patients and severe combined immunodeficiency patients undergoing retroviral gene therapy. Tightly controlled Lmo2 expression is therefore essential, yet no...
The SCL/Tal-1 gene encodes a basic helix-loop-helix transcription factor with key roles in hematopoietic and neural development. SCL is
expressed in, and required for, both primitive and definitive erythropoiesis. Thus far, we have identified only one erythroid
SCL enhancer. Located 40 kb downstream of exon 1a, the +40 enhancer displays activity in...
Transcription factors are key regulators of hematopoietic stem cells (HSCs), yet the molecular mechanisms that control their expression are largely unknown. Previously, we demonstrated that expression of Scl/Tal1, a transcription factor required for the specification of HSCs, is controlled by Ets and GATA factors. Here we characterize the molecular...
Hematopoietic stem cell (HSC) development is regulated by several signaling pathways and a number of key transcription factors, which include Scl/Tal1, Runx1, and members of the Smad family. However, it remains unclear how these various determinants interact. Using a genome-wide computational screen based on the well characterized Scl +19 HSC enhan...
Hematopoietic stem cell (HSC) development is regulated by several signaling pathways and a number of key transcription factors, which include Scl/Tal1, Runx1 and members of the Smad family. However, it remains unclear how these various determinants interact. Using a genome-wide computational screen based on the well-characterized Scl +19 HSC enhanc...
L3mbtl encodes a member of the Polycomb group of proteins, which function as transcriptional repressors in large protein complexes. The Drosophila D-l(3)mbt protein is considered a tumor suppressor since its inactivation results in brain tumors. The human L3MBTL gene lies in a region of chromosome 20 frequently deleted in patients with myeloid mali...
Appropriate transcriptional regulation is critical for the biological functions of many key regulatory genes, including the stem cell leukemia (SCL) gene. As part of a systematic dissection of SCL transcriptional regulation, we have previously identified a 5,245-bp SCL +18/19 enhancer that targeted embryonic endothelium together with embryonic and...
The stem cell leukemia (SCL) gene, also known as TAL-1, encodes a basic helix-loop-helix protein that is essential for the formation of all hematopoietic lineages, including primitive
erythropoiesis. Appropriate transcriptional regulation is essential for the biological functions of SCL, and we have previously
identified five distinct enhancers whi...
The development of blood has long served as a model for mammalian cell type specification and differentiation, and yet the underlying transcriptional networks remain ill defined. Characterization of such networks will require genome-wide identification of cis-regulatory sequences and an understanding of how regulatory information is encoded in the...
Haematopoiesis has long served as a paradigm for adult stem cell systems and studies over the last 20 years have established that transcriptional control is central to the specification and subsequent differentiation of haematopoietic stem cells (HSCs). With many of the key transcription factors known, haematopoiesis provides a powerful cellular sy...
The transcription factor SCL/TAL1 is essential for haematopoiesis and vascular development in the embryo. It is also expressed in the developing skeletal and central nervous systems, but lethality of SCL null mice has precluded detailed analysis of the role of this gene in these tissues.We have previously demonstrated that the SCL +18/19 enhancer d...
The stem cell leukaemia (SCL) gene encodes a basic helix-loop-helix transcription factor with a critical role in normal haematopoiesis and angiogenesis. The SCL gene is normally expressed in haematopoietic stem cells, mast cells, megakaryocytes, endothelium and smooth muscle. Aberrant expression of the SCL gene leads to T-cell acute lymphoblastic l...