Research progress on stable gene transfection methods of Plasmodium falciparum
- Author:
LI Xiao-song
;
PAN Mao-hua
;
HUANG Ya-ming
;
YANG Zhao-qing
- Publication Type:Review
- Keywords:
Plasmodium falciparum;
stable transfection;
efficiency;
plasmid DNA;
lyse-reseal erythrocytes for transfection
- From:
China Tropical Medicine
2023;23(2):186-
- CountryChina
- Language:Chinese
-
Abstract:
Abstract: Transfection of Plasmodium falciparum is helpful to study the function of its genes, such as drug resistance. However, transgenic manipulation has been very challenging, mainly due to the high A/T base sequence structure (A+T content of about 82%) and low transfection efficiency of the Plasmodium genome. Electroporation-based transfection of Plasmodium falciparum has been successfully applied in the study of certain genes, and electroporation by preloading is currently the preferred method for introducing foreign DNA into Plasmodium falciparum. The site-directed editing of Plasmodium genes mostly adopts the method of two-plasmid transfection. It is generally believed that successful transfection of Plasmodium requires a large amount of high-purity plasmid DNA and an accurate transfection system. In addition to the evaluation of the current commonly used electrotransfection methods, this paper also introduces a new transfection method, namely lyse-reseal erythrocytes for transfection (LyRET). This paper also review the role of factors such as plasmid DNA concentration, the use of transfection reagents, the setting of transfection parameters, the addition of fresh red blood cells, and the markers of successful transfection in improving the success rate and efficiency of Plasmodium transfection, in the hope of providing a reference for study in this field.
- Full text:16.Research progress on stable gene transfection methods of Plasmodium falciparum.pdf
