Genome Editing of the Octoploid Fragaria x ananassa Using the CRISPR/Cas9 System

Due to its octoploid nature, gene functional analyses in the cultivated strawberry (Fragaria × ananassa) are commonly carried out via gene silencing using self-complementary “hairpin” double-stranded RNA (RNAi) constructs. However, this system is not always as efficient as expected. First, an efficient silencing of the target gene is not always achieved, and second, its effect might not be stable after several clonal propagations of the transgenic lines. Recently, genome editing is becoming an important biotechnological tool for gene functional analysis and crop improvement, in particular since the development of the CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeat-CRISPR associated protein 9) system. To investigate the functionality of the CRISPR/Cas9 in strawberry, we designed two sgRNAs directed against two regions of the floral homeotic gene APETALA3 (AP3) in order to induce a deletion of around 200 nt. A vector containing both sgRNAs and Cas9 was used to transform leaf disks of F. × ananassa cv. Camarosa. Several independent stable transgenic lines displayed defects in stamen and fruit development, partially phenocopying that of the Arabidopsis ap3 mutants. Molecular analysis of the targeted AP3 locus indicated differences in gene editing among different transgenic lines and suggests mutations in all the possible AP3 alleles. Phenotypic analyses indicate that impaired fruit development might be caused by the lack of proper development of the anthers due to the CRISPR/Cas9 induced mutation in AP3. In summary, we show that the CRISPR/Cas9 system is a functional tool to perform genome editing in the octoploid F. × ananassa. We propose this system as an alternative to the traditional RNAi strategy to stably mutagenize a particular gene of interest for functional analyses in this species.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Targeted gene modification in Fragaria vesca mediated by CRISPR/Cas9 system

Genome editing is becoming an important biotechnological tool for gene function analysis and crop improvement, being the CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeat-CRISPR associated protein 9) system the most widely used. The natural CRISPR/Cas9 system has been reduced to two components: a single-guide RNA (sgRNA) for target recognition via RNA-DNA base pairing, which is commonly expressed using a promoter for small-RNAs (U6 promoter), and the Cas9 endonuclease for DNA cleavage (1). To validate the CRISPR/Cas9 system in strawberry plants, we designed two sgRNAs directed against the floral homeotic gene APETALA3 (sgRNA-AP3#1 and sgRNA-AP3#2). This gene was selected because ap3 mutations induce clear developmental phenotypes in which petals and stamens are missing or partially converted to sepals and carpels respectively (2). In this work, we used two different U6 promoters to drive the sgRNA-AP3s expression: AtU6-26 from Arabidopsis (4), and a U6 promoter from Fragaria vesca (FvU6) (this work). We also tested two different coding sequences of Cas9: a human- (hSpCas9) (3) and a plant-codon optimized (pSpCas9) (this work). Transient expression experiments using both CRISPR/Cas9 systems (AtU6-26:sgRNA-AP3#1_35S:hSpCas9_AtU6-26:sgRNA-AP3#2 and FvU6:sgRNA-AP3#1_35S:pSpCas9_FvU6:sgRNA-AP3#2) were performed infiltrating Agrobacterium tumefaciens into F. vesca fruits. PCR amplification and sequencing analyses across the target sites showed a deletion of 188-189 bp corresponding to the region comprised between the two cutting sites of Cas9, confirming that the CRISPR/Cas9 system is functional in F. vesca. Remarkably, the two systems showed different mutagenic efficiency that could be related to differences in expression of the U6 promoters as well as differences in the Cas9 transcripts stability and translation. Stable transformants for both F. vesca (2n) and Fragaria X anannassa (8n) are currently being established to test whether is possible to obtain heritable homozygous mutants derived from CRISPR/Cas9 strategies in strawberry. Thus, our work offers a promising tool for genome editing and gene functional analysis in strawberry. This tool might represent a more efficient alternative to the sometimes inefficient RNAi silencing methods commonly used in this species.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Importancia de la información para el paciente con enfermedad inflamatoria intestinal

El término enfermedad inflamatoria intestinal (EII) se emplea de forma general para referirse a enfermedades inflamatorias crónicas del intestino con un curso recurrente en forma de brotes. Generalmente al hablar de EII nos estamos refiriendo a colitis ulcerosa (CU) y enfermedad de Crohn (EC). Ambas patologías tienen una etiología aún no perfectamente conocida, pero se conoce la influencia de factores genéticos y ambientales como el tabaco. No existen evidencias firmes que asocien la EII con hábitos dietéticos o microorganismos, aunque estudios recientes relacionan la EII con características de la microbiota intestinal. Por sus características, la EII conlleva un gran impacto físico, psicológico y social que merma la calidad de vida del paciente. Por ello es vital la relación y comunicación médico-paciente, la cual contribuye a la mejora de los resultados en su salud. Objetivos: conocer el nivel de información subjetivo del paciente y sus preocupaciones; su impacto en la calidad de vida, la evolución y el tratamiento de la EII; y las principales fuentes de información del paciente.Grado en Medicin

L'importance de la visibilité dans les processus migratoires

La question de la visibilité n'a été traitée, jusqu'à présent, que de manière marginale par la sociologie des migrations. Avec cet article nous allons de démontrer la pertinence de cette notion comme instrument analytique dans le domaine d'étude des migrations. Pour accomplir cet objectif, nous allons utiliser une nouvelle méthodologie, en incorporant la visibilité dans le cadre du processus plus ample et complexe de la reconnaissance. Nous appliquerons ce schéma de travail, par la suite, au domaine des migrations en Espagne à fin de démontrer la validité de notre démarche.The Sociology of Migration has treated the «visibility» question in rather marginal way so far. With this paper we try to demonstrate the soundness of this concept as an analytical tool in migration studies. To this end, we will use a new methodology consisting in incorporating «visibility» to the wider and more complex «recognition» process. Finally, we will apply this approach to the migration question in Spain to further proof its validity

Traducción de los referentes culturales en el doblaje de la serie “Érase una vez... el hombre” al español

En el presente artículo analizaremos las dificultades que presenta la traducción audiovisual de productos de divulgación histórica para un público infantil y juvenil. El estudio se centra en dos elementos representativos del capítulo «La Revolución francesa» de la serie Érase una vez… el hombre en francés y en las versiones en español peninsular y de Latinoamérica. El objetivo será observar los problemas de traducción y las soluciones que las dos culturas y lenguas meta ofrecen.G.I. HUM 767 (ayudas a Grupos de Investigación de la Junta de Andalucía) / Editorial Comares (colección interlingua

Las gorras de paja de Bohoyo

En número dedicado a: La provincia de Ávil

Metodología de investigación en estudios de usuarios

The brief review of the history of user studies set out in this paper serves as an introduction to their definition and important role in information centres, as well as to the methods available for gathering information on users. The resulting overview of the literature is followed by a discussion of the methods used to date, where general considerations give way to an analysis of the particulars, based on an initial classification of users as active or passive information providers. The main techniques along with their advantages and drawbacks, the type of information that can be obtained with each technique and the utility of such information for the various information centre departments are also described. The final section discusses scientific methodology in general, and naturally the need to use scientific method in any type of research, in this case user surveys. The advantages afforded by the implementation of such methodology in these studies are also examined

Comparing Sexist Expressions in English and Spanish: (De)-Constructing Sexism though Language

La literatura, el folclore, los diccionarios y las gramáticas han utilizado y explicado las manifestaciones lingüísticas de modo que se adapten a la idea de que lo masculino debe ser el criterio universal (Spender 1980). Frente a estos antecedentes históricos, las feministas durante los años 70 y, posteriormente, los defensores de lo políticamente correcto de los 90, comenzaron una batalla contra el sexismo en el lenguaje, proporcionando soluciones en forma de pautas, folletos, glosarios, etc (Key 1975, Maggio 1987, 1991, McMinn 1991).El presente artículo explora la afirmación: "male equals positive and normal, female equals negative and abnormal" (Lakoff 1975, 2001) y, al prestar atención a desviaciones semánticamente parciales, proporcionar una visión histórica de equivalentes masculinos y femeninos. El corpus se construirá a partir de diccionarios monolingües y bilingües, diccionarios de expresiones idiomáticas, argot y proverbios para ofrecer un estudio diacrónico del lenguaje sexista, principalmente en inglés y español. Los sinónimos de algunas palabras se extraerán de páginas web y, para medir la actualidad de algunos de los ejemplos, se utilizarán tres corpus en línea. Se analizará también el marco sociocultural de estos términos y expresiones para ver hasta qué punto han moldeado ambas sociedades y qué efectos tienen actualmente en la vida de las mujeres.Literature, folklore, dictionaries and grammars have used and explained language manifestations to conform to the idea that male should be the standard of humanity (Spender 1980). Against this historical background, feminists during the 1970s and later politically correct advocates of the 1990s started battling against sexism in language, providing solutions, in the form of guidelines, booklets, glossaries, etc (Key 1975, Maggio 1987,1991, McMinn 1991).The present article explores the value assumption: "male equals positive and normal, female equals negative and abnormal" (Lakoff 1975, 2001) and, by paying attention to semantic biased deviations, gives a historical account of male and female counterparts. The corpus will be drawn from monolingual and bilingual dictionaries, dictionaries of idiomatic expressions, slang and proverbs to offer a diachronic study of sexist language mainly in English and in Spanish. Synonyms for certain terms will be drawn from Web pages and to measure the currency of some of the examples three on-line corpora will be used. The socio-cultural framework of these terms and expressions will be studied to see to what extent they have moulded both societies and what effects they have in women's lives today