RESUMEN
Goniodysgenesis is a developmental abnormality of the anterior chamber of the eye. It is generally considered to be congenital in dogs (Canis lupus familiaris), and has been associated with glaucoma and blindness. Goniodysgenesis and early-onset glaucoma initially emerged in Border Collies in Australia in the late 1990s and have subsequently been found in this breed in Europe and the USA. The objective of the present study was to determine the genetic basis of goniodysgenesis in Border Collies. Clinical diagnosis was based on results of examinations by veterinary ophthalmologists of affected and unaffected dogs from eleven different countries. Genotyping using the Illumina high density canine single nucleotide variant genotyping chip was used to identify a candidate genetic region. There was a highly significant peak of association over chromosome 17, with a p-value of 2 × 10-13 Expression profiles and evolutionary conservation of candidate genes were assessed using public databases. Whole genome sequences of three dogs with glaucoma, three severely affected by goniodysgenesis and three unaffected dogs identified a missense variant in the olfactomedin like 3 (OLFML3) gene in all six affected animals. This was homozygous for the risk allele in all nine cases with glaucoma and 12 of 14 other severely affected animals. Of 67 reportedly unaffected animals, only one was homozygous for this variant (offspring of parents both with goniodysgenesis who were also homozygous for the variant). Analysis of pedigree information was consistent with an autosomal recessive mode of inheritance for severe goniodysgenesis (potentially leading to glaucoma) in this breed. The identification of a candidate genetic region and putative causative variant will aid breeders to reduce the frequency of goniodysgenesis and the risk of glaucoma in the Border Collie population.
Asunto(s)
Cámara Anterior/anomalías , Proteínas de la Matriz Extracelular/genética , Glaucoma/genética , Mutación Missense , Secuencia de Aminoácidos , Animales , Cámara Anterior/metabolismo , Embrión de Pollo , Enfermedades de los Perros/genética , Enfermedades de los Perros/metabolismo , Perros/anomalías , Proteínas del Ojo/genética , Femenino , Regulación de la Expresión Génica , Estudio de Asociación del Genoma Completo , Glaucoma/metabolismo , Glaucoma/veterinaria , Glicoproteínas/genética , Humanos , Masculino , Ratones , Polimorfismo de Nucleótido Simple , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
MicroRNAs (miRNAs) are short non-coding RNAs with key roles in cellular regulation. As part of the fifth edition of the Functional Annotation of Mammalian Genome (FANTOM5) project, we created an integrated expression atlas of miRNAs and their promoters by deep-sequencing 492 short RNA (sRNA) libraries, with matching Cap Analysis Gene Expression (CAGE) data, from 396 human and 47 mouse RNA samples. Promoters were identified for 1,357 human and 804 mouse miRNAs and showed strong sequence conservation between species. We also found that primary and mature miRNA expression levels were correlated, allowing us to use the primary miRNA measurements as a proxy for mature miRNA levels in a total of 1,829 human and 1,029 mouse CAGE libraries. We thus provide a broad atlas of miRNA expression and promoters in primary mammalian cells, establishing a foundation for detailed analysis of miRNA expression patterns and transcriptional control regions.
Asunto(s)
Perfilación de la Expresión Génica/métodos , MicroARNs/genética , Anotación de Secuencia Molecular , Regiones Promotoras Genéticas/genética , Animales , Células Cultivadas , Biblioteca de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Ratones , MicroARNs/metabolismoRESUMEN
In the FANTOM5 project, transcription initiation events across the human and mouse genomes were mapped at a single base-pair resolution and their frequencies were monitored by CAGE (Cap Analysis of Gene Expression) coupled with single-molecule sequencing. Approximately three thousands of samples, consisting of a variety of primary cells, tissues, cell lines, and time series samples during cell activation and development, were subjected to a uniform pipeline of CAGE data production. The analysis pipeline started by measuring RNA extracts to assess their quality, and continued to CAGE library production by using a robotic or a manual workflow, single molecule sequencing, and computational processing to generate frequencies of transcription initiation. Resulting data represents the consequence of transcriptional regulation in each analyzed state of mammalian cells. Non-overlapping peaks over the CAGE profiles, approximately 200,000 and 150,000 peaks for the human and mouse genomes, were identified and annotated to provide precise location of known promoters as well as novel ones, and to quantify their activities.
Asunto(s)
Perfilación de la Expresión Génica , Genoma , Animales , Regulación de la Expresión Génica , Humanos , Ratones , Regiones Promotoras Genéticas , Especificidad de la EspecieRESUMEN
Lentiviral vectors have recently emerged as an efficient method of transgene delivery to the germline of animals. We now demonstrate that combining this efficiency with embryo splitting procedures enables the production of monozygotic twins, one of which is transgenic. We propose that this approach can be used to generate animals in which cell or tissue transplantation can be achieved without the use of immunosuppressive regimes.
