scholarly journals Genetic Annotation of Gain-Of-Function Screens Using RNA Interference and in Situ Hybridization of Candidate Genes in the Drosophila Wing

Genetics ◽  
2012 ◽  
Vol 192 (2) ◽  
pp. 741-752 ◽  
Author(s):  
Cristina Molnar ◽  
Mar Casado ◽  
Ana López-Varea ◽  
Cristina Cruz ◽  
Jose F. de Celis
Keyword(s):  
In Situ Hybridization ◽  
Rna Interference ◽  
Candidate Genes ◽  
Gain Of Function ◽  
Drosophila Wing

scholarly journals Exome Sequencing Implicates SWI/SNF Chromatin Remodeling Genes in Human Congenital Hydrocephalus

Neurosurgery ◽  
2019 ◽  
Vol 66 (Supplement_1) ◽  
Author(s):  
August A Allocco ◽  
Sheng Chih Jin ◽  
Weilai Dong ◽  
Xue Zeng ◽  
Sierra B Conine ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Brain Development ◽  
Candidate Genes ◽  
Chromatin Remodeling ◽  
Congenital Hydrocephalus ◽  
Normal Brain ◽  
Csf Flow ◽  
Neurodevelopmental Outcomes ◽  
Xenopus Embryos

Abstract INTRODUCTION Congenital Hydrocephalus (CH) affects 1 in 1000 live births and is treated with lifelong surgical cerebrospinal fluid (CSF) diversion with substantial morbidity. Our group recently sequenced 180 probands with CH and identified 4 novel CH genes, all of which regulate neural stem cell (NSC) fate. Nonetheless, these genes account for just 10% of studied cases. METHODS We doubled the size of our cohort through robust recruitment and exome-sequenced 361 CH probands, including 216 case-parent trios. To study the effect of novel mutations on brain development, we developed a Xenopus tropicalis model of CH using CRISPR/Cas9. Multiple, nonoverlapping CRISPR guide RNAs targeted against candidate genes were designed and injected into fertilized single cell Xenopus embryos via microinjection. Ventricular morphology and CSF flow dynamics were analyzed via optical coherence tomography (OCT) imaging. Histological sections from WT and mutant Xenopus embryos were subjected to whole-mount in Situ hybridization to assess expression of various markers of NSC growth and proliferation. RESULTS We identified new causative mutations in both previously identified and novel genes, all of which regulate ventricular zone NSC fate. Of these, enrichment in rare, damaging variants was highest in the SWI/SNF chromatin remodeling complex genes SMARCC1 and ARID1B (P = 1.83 × 10-9). Xenopus tadpoles harboring mutations in SMARCC1 exhibited marked ventriculomegaly, aqueductal stenosis, and impaired CSF flow. In Situ hybridization of SMARCC1 knockout brain sections demonstrated decreased expression of genes regulating NSC growth and proliferation. Notably, injection of wild type SMARCC1 RNA restored normal brain development in SMARCC1 KO embryos. CONCLUSION These findings implicate SWI/SNF complex genes in CH and validate the use of Xenopus for investigating additional candidate genes. These findings highlight the importance of NSC dysregulation in CH pathology, and suggest that in a subset of patients the risk of adverse neurodevelopmental outcomes may be unaltered whether or not shunting is performed.

Identifying new candidate genes for hereditary facial paresis on chromosome 3q21–q22 by RNA in situ hybridization in mouse

Genomics ◽  
2005 ◽  
Vol 86 (1) ◽  
pp. 55-67 ◽  
Author(s):  
Bert van der Zwaag ◽  
J. Peter H. Burbach ◽  
Curt Scharfe ◽  
Peter J. Oefner ◽  
Han G. Brunner ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Candidate Genes ◽  
Facial Paresis ◽  
Rna In Situ Hybridization

scholarly journals Williams-Beuren syndrome: Report of two cases with molecular diagnosis

2011 ◽  
pp. 523-528
Author(s):  
Carolina Vargas ◽  
Wilmar Saldarriaga ◽  
Harry Mauricio Pachajoa ◽  
Carolina Isaza
Keyword(s):  
Mental Retardation ◽  
In Situ Hybridization ◽  
Aortic Stenosis ◽  
Candidate Genes ◽  
Molecular Diagnosis ◽  
Fluorescent In Situ Hybridization ◽  
Chromosome 7 ◽  
Supravalvular Aortic Stenosis ◽  
Syndrome Report

Williams-Beuren syndrome has an estimated incidence of one case in every 10,000 births. It is secondary to microdeletion of a fragment in the long arm of chromosome 7, which contains several candidate genes for the characteristic phenotype of typical facies, supravalvular aortic stenosis, and variable mental retardation with a friendly personality. This article focuses on the report of two cases, with classic but varied phenotypic findings, of this syndrome for which molecular diagnosis with fluorescent in situ hybridization was available. Additionally, we suggest a protocol for complementary studies needed to characterize each patient.

Finding Candidate Genes by Preparative in Situ Hybridization

1994 ◽  
pp. 123-138
Author(s):  
J. C. Hozier ◽  
L. M. Davis ◽  
P. D. Siebert ◽  
K. Dietrich ◽  
M. C. Paterson
Keyword(s):  
In Situ Hybridization ◽  
Candidate Genes

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

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