Engineering threshold-based selection systems

Katherine H Pedone; Vanessa González-Pérez; Luciana E Leopold; Neal R Rasmussen; Channing J Der; Adrienne D Cox; Shawn Ahmed; David J Reiner

doi:10.1093/g3journal/jkab234

Engineering threshold-based selection systems

G3 (Bethesda). 2021 Sep 6;11(9):jkab234. doi: 10.1093/g3journal/jkab234.

Authors

Katherine H Pedone¹, Vanessa González-Pérez², Luciana E Leopold^{2

3}, Neal R Rasmussen⁴, Channing J Der^{1

2

5}, Adrienne D Cox^{1

2

5}, Shawn Ahmed^{2

3}, David J Reiner^{1

4

5

6}

Affiliations

¹ Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599, USA.
² Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, NC 27599, USA.
³ Department of Biology, University of North Carolina, Chapel Hill, NC 27599, USA.
⁴ Institute of Biosciences and Technology, College of Medicine, Texas A&M Health Science Center, Houston, TX 77030, USA.
⁵ Department of Pharmacology, University of North Carolina, Chapel Hill, NC 27599, USA.
⁶ Department of Radiation Oncology, University of North Carolina, Chapel Hill, NC 27599, USA.

Abstract

Using model organisms to identify novel therapeutic targets is frequently constrained by pre-existing genetic toolkits. To expedite positive selection for identification of novel downstream effectors, we engineered conditional expression of activated CED-10/Rac to disrupt Caenorhabditis elegans embryonic morphogenesis, titrated to 100% lethality. The strategy of engineering thresholds for positive selection using experimental animals was validated with pharmacological and genetic suppression and is generalizable to diverse molecular processes and experimental systems.

Keywords: 3ʹUTR; BH3-only; EGL-1; EHT 1864; EHT 8560; NMD; Pak; SMG-1; nonsense-mediated decay; small GTPase.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Caenorhabditis elegans / genetics
Caenorhabditis elegans Proteins*

Substances

Caenorhabditis elegans Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding