Abstract

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 .

Details

Title
Plate-based transfection and culturing technique for genetic manipulation of Plasmodium falciparum
Author
Caro, Florence; Miller, Mathew G; DeRisi, Joseph L
Pages
22
Publication year
2012
Publication date
2012
Publisher
BioMed Central
e-ISSN
14752875
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
926191890
Copyright
© 2012 Caro et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.