title: 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 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.
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, California, USA.
authors.forename: Florence
authors.initials: F
authors.lastname: Caro
authors.forename: Mathew G
authors.initials: MG
authors.lastname: Miller
authors.forename: Joseph L
authors.initials: JL
authors.lastname: DeRisi
mesh_terms: D005827:Genetics, Microbial / Q000379:methods*; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D010963:Plasmodium falciparum / Q000235:genetics*; D014162:Transfection / Q000379:methods*