TY - JOUR

T1 - Stable transformation of pleomorphic bloodstream form Trypanosoma brucei.

AU - MacGregor, P

AU - Rojas, F

AU - Dean, S

AU - Matthews, KR

PY - 2013/8/31

Y1 - 2013/8/31

N2 - African trypanosomes differentiate between various developmental stages both in mammalian hosts and their tsetse vector to adapt to and survive in the different environments they encounter. In the bloodstream, trypanosomes naturally exist as either proliferative slender-forms or non-proliferative stumpy-forms, the latter being responsible for both prolonged infection and transmission. However, most trypanosome studies are carried out on laboratory-adapted monomorphic cell lines, incapable of differentiating to stumpy-forms or completing the life cycle through the tsetse fly. Partly, this has been due to the inefficiency of transfection of pleomorphic strains which have retained the ability to generate stumpy-forms. Recently, Amaxa Nucleofector® technology was shown to increase transfection efficiency for monomorphic bloodstream forms. Using this technology we have optimised a similar method for pleomorphic bloodstream form transfection, generating transfection efficiencies of 10−7–10−6. This permits routine genetic manipulation of pleomorphic lines, which have the most biological relevance for trypanosomes in the field.

AB - African trypanosomes differentiate between various developmental stages both in mammalian hosts and their tsetse vector to adapt to and survive in the different environments they encounter. In the bloodstream, trypanosomes naturally exist as either proliferative slender-forms or non-proliferative stumpy-forms, the latter being responsible for both prolonged infection and transmission. However, most trypanosome studies are carried out on laboratory-adapted monomorphic cell lines, incapable of differentiating to stumpy-forms or completing the life cycle through the tsetse fly. Partly, this has been due to the inefficiency of transfection of pleomorphic strains which have retained the ability to generate stumpy-forms. Recently, Amaxa Nucleofector® technology was shown to increase transfection efficiency for monomorphic bloodstream forms. Using this technology we have optimised a similar method for pleomorphic bloodstream form transfection, generating transfection efficiencies of 10−7–10−6. This permits routine genetic manipulation of pleomorphic lines, which have the most biological relevance for trypanosomes in the field.

KW - Trypanosome

KW - Transfection

KW - Pleomorphic

KW - Stumpy

KW - Trypanosoma brucei

U2 - 10.1016/j.molbiopara.2013.06.007

DO - 10.1016/j.molbiopara.2013.06.007

M3 - Journal article

C2 - 23835071

VL - 190

SP - 60

EP - 62

JO - Molecular and Biochemical Parasitology

JF - Molecular and Biochemical Parasitology

SN - 0166-6851

IS - 2

ER -