The minimal essential host cell components for Leishmania intracellular survival: Dual RNA-seq of enucleated fibroblasts that host Leishmania amazonensis

Enucleated cells or cytoplasts (cells which nucleus is removed in vitro) represent an unexplored biologic model for intracellular infection studies due the abrupt interruption of nuclear processing and new RNA synthesis of the host cell machinery in response to pathogen entry. Using enucleated fibroblasts as host cells for the protozoan parasite Leishmania amazonensis, we demonstrate that parasite multiplication and biogenesis of large parasitophorous vacuoles are independent of host cell nucleus throughout 48 hours of intracellular parasitism. RNA sequencing of both host cytoplast and parasite transcripts (dual RNA-seq) was performed to identify which host transcripts are more conserved or degraded upon interaction with parasites and also to explore the parasite genes differentially expressed when hosted by nucleated or enucleated cells.