Departamento de Ingeniería Biomédica

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https://hdl.handle.net/1992/52212
Encuentre en acceso abierto la producción académica, investigativa y de creación del Departamento de Ingeniería Biomédica de la Universidad de los Andes.

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Examinando Departamento de Ingeniería Biomédica por Autor "52699075"

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    Evaluating CRISPR-Cas9 efficiency in the generation of nlgn-1 knock-out in one-cell stage zebrafish embryos as a model for the study of stress behavior
    (Universidad de los Andes, 2022-12-12) Bonilla Paez, Andrea del Pilar; Bloch Morel, Natasha Ivonne; Akle Álvarez, Verónica; 52863778; 52699075; Muñoz Camargo, Carolina; Lattig Matiz, María Claudia; Neurociencia Traslacional
    Neuroligins (NL) are transsynaptic cell adhesion proteins involved in shaping synapsis when they interact with presynaptic neurexins proteins. Evidence continues to accumulate on their association with some neuropsychiatric disorders making them relevant proteins of study. The neuroligin-1 gene (nlgn1), a member of the NL family, involved in excitatory synapses, has been associated with the genetic bases of stress-related disorders (major depression disorder and PTSD). However, its specific role in the mediation of stress has not been investigated in detail. Zebrafish (Danio rerio) has emerged as a model in translational neuroscience and behavioral research, that allows the study of behaviors related to anxiety, fear, sleep, stress, and memory, making it an ideal model for such a study. Here, we aim to generate a nlgn1-crispant zebrafish using a CRISPR-Cas9 system and evaluate the effect of this genetic modification on larval behavior. For this purpose, we had to standardize the CRISPR gene editing system. We initially generated tyr-crispant zebrafish embryos using CRISPR-cas9 as this produces a visible phenotype. We verified our CRISPR gene editing system with tyr, produced crispant-like phenotypes with varying degrees of mosaicism with a ~76% efficiency. Then, zebrafish embryos injected with sgRNAs targeting nlgn1 gene showed a low rate of survival rates between 11.9% and 7 % depending on the sgRNA used. Moreover, many of the embryos showed malformations during development that became lethal at 3dpf. Differences in immobility mean during open field test were significantly different between nlgn1-crispant and controls. These results suggest that neuroligin-1 might have a crucial role during development or a strong neurological phenotype.