TY  - GEN
T1  - Unlike Drosophila elav, the C. elegans elav orthologue exc-7 is not panneuronally expressed
AU  - Pham, Kenneth
AU  - Hobert, Oliver
DO  - 10.17912/micropub.biology.000189
UR  - http://beta.micropublication.org/journals/biology/micropub-biology-000189/
AB  - We are interested in identifying genes that are expressed in a panneuronal manner throughout the nervous system (Stefanakis et al., 2015). The Drosophila elav locus is a panneuronally expressed RNA binding protein (Campos et al., 1987; Robinow and White, 1988). Elav protein staining is routinely used in Drosophila to identify neurons and cis-regulatory control regions from the elav locus are routinely used as panneuronal Gal4 drivers (Berger et al., 2007; Luo et al., 1994; O’Neill et al., 1994; Osterwalder et al., 2001; Robinow and White, 1991). Based on sequence homology, the C. elegans exc-7 locus is the sole C. elegans orthologue of elav (Fujita et al., 2003; Fujita et al., 1999; Loria et al., 2003; Samson, 2008). Previous expression pattern analyses have shown that exc-7 is expressed only in a subset of neurons of the nervous system (the expressing neurons were mostly unidentified) (Fujita et al., 1999; Loria et al., 2003). However, reporter gene constructs previously used to infer exc-7 expression did not contain all intergenic region of the large exc-7 locus. Therefore, the possibility remained that through the use of more distal cis-regulatory elements, C. elegans exc-7 could also be panneuronally expressed, like its fly orthologue. To address this possibility, we tagged endogenous exc-7 with gfp::3xflag at its C-terminus using CRISPR/Cas9 genome engineering (Dokshin et al., 2018) and examined its expression. We cloned gfp from the che-1(ot856[che-1::gfp]) allele (Leyva-Diaz and Hobert, 2019), inserted it into the pMiniT 2.0 vector (NEB), and used that resulting plasmid for subsequent cloning of the gfp tag.
PY  - 2019
JO  - microPublication Biology
ER  -