A simple strategy for addition of degron tags to endogenous genes harboring prior insertions of fluorescent protein. Academic Article uri icon

abstract

  • There exist insufficient validated "entry portal" sites in the C. elegans genome for CRISPR/Cas9-dependent insertion into endogenous genes to confer diverse spatiotemporal patterns and levels of expression on exogenous sequences. Consequently, we recognized the most common potential "entry portal" sequences: genes previously tagged with fluorescent proteins using CRISPR/Cas9. As proof of concept, we used existing mKate2-encoding sequences inserted in the 5' end of genes as an insertion point for the auxin inducible degron, AID*. This sequence permits reasonably efficient insertion that can be employed using a variety of approaches for different end goals. Our strategy is thus generalizable to many needs.

published proceedings

  • MicroPubl Biol

author list (cited authors)

  • Fakieh, R., Duong, T., Wu, Y., Rasmussen, N., & Reiner, D.

complete list of authors

  • Fakieh, Razan||Duong, Tam||Wu, You||Rasmussen, Neal||Reiner, David

publication date

  • August 2022