Deep mutational scanning of the RNase III-like domain in Trypanosoma brucei RNA editing protein KREPB4.

ABSTRACT: Kinetoplastid pathogens including Trypanosoma brucei, T. cruzi, and Leishmania species, are early diverged, eukaryotic, unicellular parasites. Functional understanding of many proteins from these pathogens has been hampered by limited sequence homology to proteins from other model organisms. Here we describe the development of a high-throughput deep mutational scanning approach in T. brucei that facilitates rapid and unbiased assessment of the impacts of many possible amino acid substitutions within a protein on cell fitness, as measured by relative cell growth. The approach leverages several molecular technologies: cells with conditional expression of a wild-type gene of interest and constitutive expression of a library of mutant variants, degron-controlled sta

SUBMITTER: McDermott SM 

PROVIDER: S-EPMC11033214 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature