Plate-based transfection and culturing technique for genetic manipulation of Plasmodium falciparum

Abstract Genetic manipulation of malaria parasites remains an inefficient, time-consuming and resource-intensive process. Presented here is a set of methods for 96-well plate-based transfection and culture that improve the efficiency of genetic manipulation of Plasmodium falciparum . Compared to sta...

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Bibliographic Details
Published in:Malaria Journal
Main Authors: Caro Florence, Miller Mathew G, DeRisi Joseph L
Format: Article in Journal/Newspaper
Language:English
Published: BMC 2012
Subjects:
malaria
Plasmodium falciparum
transfection
96-well plate
Arctic medicine. Tropical medicine
RC955-962
Infectious and parasitic diseases
RC109-216
Arctic
Online Access:https://doi.org/10.1186/1475-2875-11-22
https://doaj.org/article/46d4621c27d14f6195da9d7bbea270c3
Description
Summary:Abstract Genetic manipulation of malaria parasites remains an inefficient, time-consuming and resource-intensive process. Presented here is a set of methods for 96-well plate-based transfection and culture that improve the efficiency of genetic manipulation of Plasmodium falciparum . Compared to standard protocols plate-based transfection requires 20-fold less DNA, transient transfection efficiency achieved is approximately seven-fold higher, whilst stable transfection success rate is above 90%. Furthermore the utility of this set of protocols to generate a knockout of the PfRH3 pseudogene, screened by whole-cell PCR, is demonstrated. The methods and tools presented here will facilitate genome-scale genetic manipulation of P. falciparum .

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