Creation of a novel CRISPR-generated allele to express HA epitope-tagged C1QL1 and improved methods for its detection at synapses.

Autor: Cheung HW; Department of Neuroscience, University of Connecticut Health, Farmington, CT, USA., Schouw AD; Department of Neuroscience, University of Connecticut Health, Farmington, CT, USA., Altunay ZM; Department of Neuroscience, University of Connecticut Health, Farmington, CT, USA., Maddox JW; Department of Neuroscience, University of Texas-Austin, TX, USA., Kresic LC; Department of Neuroscience, University of Connecticut Health, Farmington, CT, USA., McAllister BC; Department of Neuroscience, University of Connecticut Health, Farmington, CT, USA., Caro K; Department of Neuroscience, University of Connecticut Health, Farmington, CT, USA., Alam S; Department of Neuroscience, University of Connecticut Health, Farmington, CT, USA., Huang A; Department of Neuroscience, University of Texas-Austin, TX, USA., Pijewski RS; Department of Neuroscience, University of Connecticut Health, Farmington, CT, USA.; Department of Biology, Anna Maria College, Paxton, MA, USA., Lee A; Department of Neuroscience, University of Texas-Austin, TX, USA., Martinelli DC; Department of Neuroscience, University of Connecticut Health, Farmington, CT, USA.; The Connecticut Institute for the Brain and Cognitive Sciences (IBACS), Storrs, CT, USA.
Jazyk: angličtina
Zdroj: FEBS letters [FEBS Lett] 2024 Oct; Vol. 598 (19), pp. 2417-2437. Date of Electronic Publication: 2024 Jun 10.
DOI: 10.1002/1873-3468.14946
Abstrakt: C1QL1 is expressed in a subset of cells in the brain and likely has pleiotropic functions, including the regulation of neuron-to-neuron synapses. Research progress on C1QL proteins has been slowed by a dearth of available antibodies. Therefore, we created a novel knock-in mouse line in which an HA-tag is inserted into the endogenous C1ql1 locus. We examined the entire brain, identifying previously unappreciated nuclei expressing C1QL1, presumably in neurons. By total numbers, however, the large majority of C1QL1-expressing cells are of the oligodendrocyte lineage. Subcellular immunolocalization of synaptic cleft proteins is challenging, so we developed a new protocol to improve signal at synapses. Lastly, we compared various anti-HA antibodies to assist future investigations using this and likely other HA epitope-tagged alleles.
(© 2024 Federation of European Biochemical Societies.)
Databáze: MEDLINE
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