Application of CRISPR/Cas9 to the study of brain development and neuropsychiatric disease

Powell, Samuel K.; Gregory, Jill; Akbarian, Schahram; Brennand, Kristen J.

doi:10.1016/j.mcn.2017.05.007

Cited by 27 publications

(14 citation statements)

References 143 publications

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“…Personalized medicine is a rapidly progressing field with immense future potential in both diagnostics and therapeutics, especially given the recent advances in CRISPR/Cas9 genome‐editing technology . Identifying molecular and genetic biomarkers will have an impact on the field of cerebrovascular medicine.…”

Section: Discussionmentioning

confidence: 99%

Polymorphism in dural arteriovenous fistula: matrix metalloproteinase‐2‐1306 C/T as a potential risk factor for sinus thrombosis

Tsuei

Chou

Chen

et al. 2018

Journal of Thrombosis and Haemostasis

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Background Dural arteriovenous fistula (DAVF) is a rare but important cerebrovascular disorder in adults. Little is known about the molecular genetic pathogenesis underlying DAVF development. Objectives To investigate the associations of gene polymorphisms and DAVF. Materials and Methods By the use of real-time PCR genotyping, seven single-nucleotide polymorphisms (SNPs) of angiogenesis-related genes were analyzed in 72 DAVF patients. Pertinent clinical and imaging data were subgrouped on the basis of location (cavernous sinus versus lateral sinus), lesions (single versus multiple), cerebral venous reflux (CVR) grading (Borden I versus Borden II/III), and sinus thrombosis (with versus without). Results We found that individuals carrying the polymorphic allele of matrix metalloproteinase (MMP)-2-1306 C/T (rs243865) had a significantly increased risk of sinus thrombosis in DAVF (odds ratio 6.2; 95% confidence interval 1.7-22.9). There was a weak difference in associations of tissue inhibitor of metalloproteinase (TIMP)-2 (rs2277698) gene polymorphism and DAVF patients subgrouped by CVR grading. Conclusions These preliminary results indicate that MMP-2-1306 C/T, but not MMP-9, TIMP-1, TIMP-2, and vascular endothelial growth factor A SNP variants, is a risk factor for the development of sinus thrombosis in DAVF patients.

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Section: Discussionmentioning

confidence: 99%

Polymorphism in dural arteriovenous fistula: matrix metalloproteinase‐2‐1306 C/T as a potential risk factor for sinus thrombosis

Tsuei

Chou

Chen

et al. 2018

Journal of Thrombosis and Haemostasis

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“…The CRISPR/Cas9 system can be used in a variety of ways to study neurological and psychiatric diseases in stem cell-based models ( 70 , 71 ). One particularly promising application is the manipulation of genetic material to explore the role of putative allelic variations in neurological and psychiatric conditions.…”

Section: Methods and Advances In Generating Hipsc-derived Neural Progmentioning

confidence: 99%

Modeling Neuropsychiatric and Neurodegenerative Diseases With Induced Pluripotent Stem Cells

et al. 2018

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Human-induced pluripotent stem cells (hiPSCs) have revolutionized our ability to model neuropsychiatric and neurodegenerative diseases, and recent progress in the field is paving the way for improved therapeutics. In this review, we discuss major advances in generating hiPSC-derived neural cells and cutting-edge techniques that are transforming hiPSC technology, such as three-dimensional “mini-brains” and clustered, regularly interspersed short palindromic repeats (CRISPR)-Cas systems. We examine specific examples of how hiPSC-derived neural cells are being used to uncover the pathophysiology of schizophrenia and Parkinson’s disease, and consider the future of this groundbreaking research.

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“…We hypothesized that C11orf46's affinity to the KMT-RC could be exploited, in order to 'edit' neuronal gene expression via a chromatin-associated mechanism. Such type of epigenome editing could be accomplished, for example, by RNA-guided fusion proteins comprised of nuclease-deficient clustered, regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein Cas9 (dCas9) bound to a transcriptional activator or repressor 38 .…”

Section: Supplementary Fig 3 and Supplementarymentioning

confidence: 99%

“…However, because '1:1' systems (one transcriptional regulator per dCas9 copy) are often only minimally effective 38,39 , we instead used dCas9-SunTag protein scaffolds which assembles as a binary system with the protein of interest fused to a single-chain antibody variable fragment (scFv)-green fluorescent protein cassette, with the scFv binding to the Cas9-fused protein scaffold carrying 10 or more copies of GCN4 (the scFv epitope) 39 . We built a dCas9-SunTag system to load ten copies of C11orf46, or the VP64 activator protein as a positive control, onto a single sgRNA-target sequence (Fig.…”

Section: Supplementary Fig 3 and Supplementarymentioning

confidence: 99%

In vivo epigenetic editing of sema6a promoter reverses impaired transcallosal connectivity caused by C11orf46/ARL14EP neurodevelopmental risk gene

Peter

Saito²,

Hasegawa

et al. 2018

Preprint

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Many neuropsychiatric risk genes contribute to epigenetic regulation of gene expression but very little is known about specific chromatin-associated mechanisms governing the formation and maintenance of neuronal connectivity. Here we show that transcallosal connectivity is critically dependent on C11orf46 (also known as ARL14EP), a small nuclear protein encoded in the chromosome 11p13 Wilms Tumor, Aniridia, Genitourinary Abnormalities, intellectual disability (formerly referred to as Mental Retardation) (WAGR) risk locus.C11orf46 haploinsufficiency in WAGR microdeletion cases was associated with severe hypoplasia of the corpus callosum. In utero short hairpin RNA-mediated C11orf46 knockdown disrupted transcallosal projections of cortical pyramidal neurons, a phenotype that was rescued by wild type C11orf46 but not the C11orf46 R236H mutant associated with autosomal recessive intellectual disability. Multiple genes encoding key regulators of axonal growth and differentiation, including Sema6A, were hyperexpressed in C11orf46-knockdown neurons.Importantly, RNA-guided epigenetic editing of neuronal Sema6a gene promoters via a dCas9 proteinconjugated SunTag scaffold with multimeric (10x) C11orf46 binding during early developmental periods, resulted in normalization of expression and rescue of transcallosal dysconnectivity via repressive chromatin remodeling, including up-regulated histone H3K9 methylation by the KAP1-SETDB1 repressor complex. Our study demonstrates that interhemispheric communication is highly sensitive to locus-specific remodeling of neuronal chromatin, revealing the therapeutic potential for shaping the brain's connectome via gene-targeted designer activators and repressor proteins. the FOXG1 transcription factor 12 . However, molecular and cellular mechanisms linking the neuronal epigenome to interhemispheric connectivity remain poorly explored.Here, we describe C11orf46/ARL14EP, a small (35kDa) nuclear protein encoded in the chr. 11p13 Wilms Tumor, Aniridia, Genitourinary Abnormalities, intellectual disability (formerly referred to as Mental Retardation) (WAGR) risk locus, as a novel chromatin regulator of transcallosal connectivity. Importantly, weshow that C11orf46 is, as an RNA-guided Cas9 fusion protein, suitable for targeted epigenomic promoter editing to drive the expression of specific regulators of axonal development in immature transcallosal projection neurons, thereby offering a molecular tool to affect interhemispheric communication. ResultsC11orf46 is expressed in cortical projection neurons and critical for transcallosal connectivity. A recent study identified 50 novel genes associated with autosomal recessive forms of intellectual disability, including open reading frame C11orf46 13 . Importantly, moreover, this gene is located in the chr. 11p13-14 neurodevelopmental risk locus and deleted in some cases of WAGR syndrome (Online Mendelian Inheritance of Man (OMIM) #194072), a rare copy number variant disorder with its core features caused by haploinsufficiency for the Wilms Tumor 1 (...

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Application of CRISPR/Cas9 to the study of brain development and neuropsychiatric disease

Cited by 27 publications

References 143 publications

Polymorphism in dural arteriovenous fistula: matrix metalloproteinase‐2‐1306 C/T as a potential risk factor for sinus thrombosis

Polymorphism in dural arteriovenous fistula: matrix metalloproteinase‐2‐1306 C/T as a potential risk factor for sinus thrombosis

Modeling Neuropsychiatric and Neurodegenerative Diseases With Induced Pluripotent Stem Cells

In vivo epigenetic editing of sema6a promoter reverses impaired transcallosal connectivity caused by C11orf46/ARL14EP neurodevelopmental risk gene

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