Silva-Garcia CG et al. (2019) G3 (Bethesda) "Single-Copy Knock-In Loci for Defined Gene Expression in Caenorhabditis elegans."

Dutta S, Heintz C, Mair WB, Silva-Garcia CG, Lanjuin A, Clark NM

[G3 (Bethesda), 2019]

We have generated single-copy knock-in loci for defined gene expression (SKI LODGE) system to insert any DNA by CRISPR/Cas9 at defined safe harbors in the <i>Caenorhabditis elegans</i> genome. Utilizing a single crRNA guide, which also acts as a Co-CRISPR enrichment marker, any DNA sequence can be introduced as a single copy, regulated by different tissue-specific promoters. The SKI LODGE system provides a fast, economical and effective approach for generating single-copy ectopic transgenes in <i>C. elegans</i>.

Fitzgerald K et al. (2006) PLoS Genet "Chemical genetics reveals an RGS/G-protein role in the action of a compound."

Ross-Macdonald P, Burley B, Thalody G, Fitzgerald K, van der Linden A, Bjerke L, Doberstein S, Kindt RM, Kopczynski J, Walker SG, Starrett J, Zhang Y, Korswagen H, Plasterk R, Choy R, Cockett M, Lodge NJ, Franke Y, Wayne H, Krystek SR, Wardwell-Swanson J, Stouch T, Dubaquie Y, Carroll P, Cao J, Tertyshnikova S, Moore L

[PLoS Genet, 2006]

We report here on a chemical genetic screen designed to address the mechanism of action of a small molecule. Small molecules that were active in models of urinary incontinence were tested on the nematode Caenorhabditis elegans, and the resulting phenotypes were used as readouts in a genetic screen to identify possible molecular targets. The mutations giving resistance to compound were found to affect members of the RGS protein/G-protein complex. Studies in mammalian systems confirmed that the small molecules inhibit muscarinic G-protein coupled receptor (GPCR) signaling involving G-alphaq (G-protein alpha subunit). Our studies suggest that the small molecules act at the level of the RGS/G-alphaq signaling complex, and define new mutations in both RGS and G-alphaq, including a unique hypo-adapation allele of G-alphaq. These findings suggest that therapeutics targeted to downstream components of GPCR signaling may be effective for treatment of diseases involving inappropriate receptor activation.