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

Authors :
Cheung, Hiu W.
Schouw, Alexander D.
Altunay, Zeynep M.
Maddox, J. Wesley
Kresic, Lyndsay C.
McAllister, Brenna C.
Caro, Keaven
Alam, Shahnawaz
Huang, Angie
Pijewski, Robert S.
Lee, Amy
Martinelli, David C.
Source :
FEBS Letters; Oct2024, Vol. 598 Issue 19, p2417-2437, 21p
Publication Year :
2024

Abstract

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. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
00145793
Volume :
598
Issue :
19
Database :
Complementary Index
Journal :
FEBS Letters
Publication Type :
Academic Journal
Accession number :
180280132
Full Text :
https://doi.org/10.1002/1873-3468.14946