Genetic manipulation of cell line derived reticulocytes enables dissection of host malaria invasion requirements

Timothy J. Satchwell*, Katherine E. Wright, Katy L. Haydn-Smith, Fernando Sánchez-Román Terán, Pedro L. Moura, Joseph Hawksworth, Jan Frayne, Ashley M. Toye, Jake Baum

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

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Abstract

Investigating the role that host erythrocyte proteins play in malaria infection is hampered by the genetic intractability of this anucleate cell. Here we report that reticulocytes derived through in vitro differentiation of an enucleation-competent immortalized erythroblast cell line (BEL-A) support both successful invasion and intracellular development of the malaria parasite Plasmodium falciparum. Using CRISPR-mediated gene knockout and subsequent complementation, we validate an essential role for the erythrocyte receptor basigin in P. falciparum invasion and demonstrate rescue of invasive susceptibility by receptor re-expression. Successful invasion of reticulocytes complemented with a truncated mutant excludes a functional role for the basigin cytoplasmic domain during invasion. Contrastingly, knockout of cyclophilin B, reported to participate in invasion and interact with basigin, did not impact invasive susceptibility of reticulocytes. These data establish the use of reticulocytes derived from immortalized erythroblasts as a powerful model system to explore hypotheses regarding host receptor requirements for P. falciparum invasion.

Original languageEnglish
Article number3806 (2019)
Number of pages9
JournalNature Communications
Volume10
DOIs
Publication statusPublished - 23 Aug 2019

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  • Student Theses

    Investigating the process of reticulocyte maturation to erythrocytes in vitro

    Author: Moura, P. L., 23 Jan 2020

    Supervisor: Toye, A. (Supervisor) & Frayne, J. (Supervisor)

    Student thesis: Doctoral ThesisDoctor of Philosophy (PhD)

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