A targeted CRISPR-Cas9 mediated F0 screen identifies genes involved in establishment of the enteric nervous system.

Autor: Moreno-Campos R; Biosciences Department, Rice University, Houston, Texas, United States of America.; Laboratory of Neural Crest and Enteric Nervous System Development, Rice University, Houston, Texas, United States of America., Singleton EW; Biosciences Department, Rice University, Houston, Texas, United States of America.; Laboratory of Neural Crest and Enteric Nervous System Development, Rice University, Houston, Texas, United States of America., Uribe RA; Biosciences Department, Rice University, Houston, Texas, United States of America.; Laboratory of Neural Crest and Enteric Nervous System Development, Rice University, Houston, Texas, United States of America.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2024 May 29; Vol. 19 (5), pp. e0303914. Date of Electronic Publication: 2024 May 29 (Print Publication: 2024).
DOI: 10.1371/journal.pone.0303914
Abstrakt: The vertebrate enteric nervous system (ENS) is a crucial network of enteric neurons and glia resident within the entire gastrointestinal tract (GI). Overseeing essential GI functions such as gut motility and water balance, the ENS serves as a pivotal bidirectional link in the gut-brain axis. During early development, the ENS is primarily derived from enteric neural crest cells (ENCCs). Disruptions to ENCC development, as seen in conditions like Hirschsprung disease (HSCR), lead to the absence of ENS in the GI, particularly in the colon. In this study, using zebrafish, we devised an in vivo F0 CRISPR-based screen employing a robust, rapid pipeline integrating single-cell RNA sequencing, CRISPR reverse genetics, and high-content imaging. Our findings unveil various genes, including those encoding opioid receptors, as possible regulators of ENS establishment. In addition, we present evidence that suggests opioid receptor involvement in the neurochemical coding of the larval ENS. In summary, our work presents a novel, efficient CRISPR screen targeting ENS development, facilitating the discovery of previously unknown genes, and increasing knowledge of nervous system construction.
Competing Interests: The authors have declared no competing interests exist.
(Copyright: © 2024 Moreno-Campos et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
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