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This essay examines Tlaquepaque's role in the consummation of New Spain's independence in 1821. Serving as a meeting center for Creole and Spanish elites, Tlaquepaque was the stage for discussions on political and economic reconfiguration. According to Barbosa-Ramírez (1981); the town became a center for social gatherings similar to Michoacán. A group of local investors and merchants sought to transform the colonial order, promoting greater autonomy and commercial openness. In this regard, Olveda (1991, 2003); highlights how these actors took advantage of the power vacuum to proclaim independence on June 13, 1821. In the same vein, Pérez (1910); documents the proclamation that challenged existing political structures and sought to redefine economic relations. Along these lines, Timothy (1991); and Zoraida (2003); argue that this event laid the foundations for federalism in the 19th century. In accordance with this, Marichal, del Valle, Kraselsky, Salvucci, and García (2019); emphasize that the movement was motivated by the economic interests of local groups seeking to expand their commercial influence. The essay, supported by these diverse historiographical perspectives, reveals the complexity of political and economic factors that shaped the independence process in this region of New Spain., Este ensayo examina el papel de Tlaquepaque en la consumación de la independencia de Nueva España en 1821. Fungiendo como centro de reunión para élites criollas y españolas, Tlaquepaque fue escenario de discusiones sobre la reconfiguración política y económica. Según Barbosa-Ramírez (1981); la villa se convirtió en un centro de tertulias similar a Michoacán. Un grupo de inversionistas y comerciantes locales buscó transformar el orden colonial, promoviendo mayor autonomía y apertura comercial. En este sentido, Olveda (1991, 2003); destaca cómo estos actores aprovecharon el vacío de poder para proclamar la independencia el 13 de junio de 1821. En este mismo orden de ideas, Pérez (1910); documenta la proclama que desafió las estructuras políticas existentes y buscó redefinir las relaciones económicas. En esa misma línea, Timothy (1991); y Zoraida (2003); argumentan que este evento sentó las bases del federalismo en el siglo XIX. Acorde con esto, Marichal, del Valle, Kraselsky, Salvucci y García (2019); subrayan que el movimiento estuvo motivado por intereses económicos de grupos locales que buscaban expandir su influencia comercial. El ensayo, respaldado por estas diversas perspectivas historiográficas, revela la complejidad de factores políticos y económicos que moldearon el proceso de independencia en esta región de Nueva España.

CRISPR-RfxCas13d system induces effective mRNA knockdown in vertebrate embryos, including zebrafish and mouse embryos. Until recently it was considered a precise tool to induce RNA depletion in an efficient and specific manner, however, this has been up to debate due to the recent discovery of collateral activity in eukaryotic cells. This phenomenon previously described in bacteria and in vitro consists in the degradation of non-target RNAs by the Cas13 protein in a non-controlled manner upon the previous specific targeting mediated by the gRNA. The need to further study the activity of this tool in different contexts has brought us to characterize the collateral activity of RfxCas13d in zebrafish embryos and to optimize other RNA-targeting CRISPR-Cas tools. Here, we demonstrate that CRISPR-RfxCas13d can be used to reliably deplete the vast majority of naturally present RNAs in zebrafish embryos without inducing collateral activity. We also discuss the potential of two other CRISPR-Cas systems, Cas7-11 and DjCas13d, as well as a high-fidelity version of RfxCas13d, as alternatives for the depletion of extremely abundant RNAs in zebrafish embryos, analysing their on-target and collateral activity. Overall, this work will contribute to enhance CRISPR-Cas approaches to target RNA in vivo, that ultimately will help to potentially use this technology as a precise therapeutic tool.

Heliports are facilities that play a fundamental role in security and emergency operations. Since rotorcrafts do not need much space for take-off and landing, heliports are normally immersed in the urban fabric of our cities. However, they generate high noise levels, which can cause a nuisance, especially in outdoor areas intended for the recreation of citizens. This paper studies how helicopter noise affects the perception of the soundscape appropriateness and landscape quality in the vicinity of a heliport located in an urban park, using semantic differential scales and appraisals on the noise sources. The study area was the "Parque del Bicentenario" in Quito, Ecuador. Immersive Virtual Reality (IVR) laboratory tests using 360-degree videos and spatial audio were preferred to on-site questionnaires, given the difficulty of predicting when helicopter noise events would occur. For the statistical analysis, objective acoustic and psychoacoustic parameters have also been considered. Results show that the soundscape is perceived as more pleasant and less chaotic when there is no helicopter noise. Furthermore, with the same visual stimuli, the appraisals of the landscape are much better in the scenarios without the helicopter noise. Sharpness is the psychoacoustic parameter that best explains the variance of the subjective variables evaluated.

The requirement for Cas nucleases to recognize a specific PAM is a major restriction for genome editing. SpCas9 variants SpG and SpRY, recognizing NGN and NRN PAMs, respectively, have contributed to increase the number of editable genomic sites in cell cultures and plants. However, their use has not been demonstrated in animals. Here we study the nuclease activity of SpG and SpRY by targeting 40 sites in zebrafish and C. elegans. Delivered as mRNA-gRNA or ribonucleoprotein (RNP) complexes, SpG and SpRY were able to induce mutations in vivo, albeit at a lower rate than SpCas9 in equivalent formulations. This lower activity was overcome by optimizing mRNA-gRNA or RNP concentration, leading to mutagenesis at regions inaccessible to SpCas9. We also found that the CRISPRscan algorithm could help to predict SpG and SpRY targets with high activity in vivo. Finally, we applied SpG and SpRY to generate knock-ins by homology-directed repair. Altogether, our results expand the CRISPR-Cas targeting genomic landscape in animals.

CRISPR-Cas systems have been used to induce DNA mutagenesis for gene function discovery. However, the development of tools to eliminate RNAs provides complementary and unique approaches to disrupt gene expression. Here, we present a workflow to perform specific, efficient, and cost-effective mRNA knockdown in zebrafish embryos using our in vivo optimized CRISPR-RfxCas13d (CasRx) system. Although the described protocol focuses on mRNA knockdown in zebrafish embryos, it can also be applied to other vertebrates.