Density sensing RIT-seq

Bloodstream-form Trypanosoma brucei maintained in culture grow to a peak cell density, which is far below that achieved by insect-stage parasites in vitro. To identify parasite factors that may act to inhibit in vitro growth at high density, we maintained our induced genome-scale BSF RNAi library at or above 1 million cells per ml for three weeks. RIT-seq identified a limited panel of proteins whose RNAi depletion supported in vitro parasite growth at high density.