AI Article Synopsis
- CRISPR/Cas9 gene knockout is a powerful method for studying gene function, particularly in animal development, but has been limited in application to specific tissues.
- Researchers introduced a new toolkit called CRISPR-TRiM to selectively knockout genes in motoneurons, muscles, and glial cells, validating its effectiveness across these tissues.
- Their findings revealed the importance of SNARE pathways in maintaining neuromuscular junctions and highlighted the role of the ESCRT pathway in controlling bouton growth and axon termini functionality in motoneurons.
Article Abstract
Tissue-specific gene knockout by CRISPR/Cas9 is a powerful approach for characterizing gene functions in animal development. However, this approach has been successfully applied in only a small number of tissues. The motor nervous system is an excellent model system for studying the biology of neuromuscular junction (NMJ). To expand tissue-specific CRISPR to the motor system, here we present a CRISPR-mediated tissue-restricted mutagenesis (CRISPR-TRiM) toolkit for knocking out genes in motoneurons, muscles, and glial cells. We validated the efficacy of this toolkit by knocking out known genes in each tissue, demonstrated its orthogonal use with the Gal4/UAS binary expression system, and showed simultaneous knockout of multiple redundant genes. Using these tools, we discovered an essential role for SNARE pathways in NMJ maintenance. Furthermore, we demonstrate that the canonical ESCRT pathway suppresses NMJ bouton growth by downregulating the retrograde Gbb signaling. Lastly, we found that axon termini of motoneurons rely on ESCRT-mediated intra-axonal membrane trafficking to lease extracellular vesicles at the NMJ.
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|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10557614 | PMC |
| http://dx.doi.org/10.1101/2023.09.25.559303 | DOI Listing |
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