In vitro selection of RNA aptamers against a conserved region of the Plasmodium falciparum erythrocyte membrane protein 1.
The var-gene encoding Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is known to play a major role in the pathogenicity of the P. falciparum parasite. The protein enables the parasite to adhere to the endothelial linings of small blood vessels (cytoadherence) as well as to non-infected erythrocytes (rosetting), thus preventing clearance from the bloodstream. The development and spread of resistance towards most anti-malarial drugs used for treatment and prevention of the most severe form of malaria truly emphasise the importance of a continuous research and development of new drugs. In this study we use Systematic Evolution of Ligands by EXponential enrichment (SELEX) methodology to isolate high-affinity ligands (aptamers). To validate the results from the SELEX in vitro selection, different aptamers have been selected against PfEMP1 in a live cell assay of P. falciparum strain FCR3S1.2, a highly rosetting strain. We have been able to show the rosette disrupting capacity of these SELEX-aptamers at concentrations of 33 nM and with 100% disruption at 387 nM. The described results show that RNA aptamers are promising candidates for adjunct therapy in severe malaria.
Barfod
Anders
A
Department of Organic Chemistry, Lund University, 22100, Lund, Sweden.
Persson
Tina
T
Lindh
Johan
J
D000818:Animals; D000962:Antimalarials / Q000302:isolation & purification*; D052157:Aptamers, Nucleotide / Q000235:genetics* / Q000302:isolation & purification*; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000235:genetics*; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000235:genetics; D016053:RNA, Protozoan / Q000037:antagonists & inhibitors / Q000235:genetics; D012397:Rosette Formation; D052156:SELEX Aptamer Technique
D000962:Antimalarials; D052157:Aptamers, Nucleotide; D015800:Protozoan Proteins; D012333:RNA, Messenger; D016053:RNA, Protozoan; C085731:erythrocyte membrane protein 1, Plasmodium falciparum
Curr Opin Infect Dis. 2005 Dec;18(6):531-6
16258328
Biomol Eng. 2007 Oct;24(4):381-403
17627883
Malar J. 2004 Dec 15;3:50
15601471
Anal Biochem. 1976 May 7;72:248-54
942051
Mol Biochem Parasitol. 2004 Mar;134(1):3-9
14747137
J Exp Med. 2000 Jul 3;192(1):1-10
10880521
Nature. 1990 Aug 30;346(6287):818-22
1697402
Vaccine. 2004 Jul 29;22(21-22):2701-12
15246600
J Biotechnol. 2000 Mar;74(1):27-38
10943570
J Biol Chem. 2002 Jun 7;277(23):20756-62
11919187
Cell. 1995 Jul 14;82(1):77-87
7541722
PLoS Pathog. 2006 Sep;2(9):e100
17009869
Br Med Bull. 1998;54(3):703-15
10326295
Vaccine. 2006 Aug 28;24(35-36):6140-54
16837110
Microbes Infect. 2007 May;9(6):777-83
17398133
Malar J. 2006 Aug 03;5:67
16887017
Proc Natl Acad Sci U S A. 2007 Oct 2;104(40):15835-40
17895392
Science. 1991 Jul 19;253(5017):314-7
1857967
Science. 1990 Aug 3;249(4968):505-10
2200121
Infect Immun. 2000 May;68(5):2971-5
10768996
Am J Physiol. 1999 Jun;276(6):C1231-42
10362584
Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
12824337
Proteomics. 2004 Apr;4(4):1086-93
15048989
J Biomol Tech. 2005 Sep;16(3):224-34
16461946
Protein Sci. 1998 Mar;7(3):706-19
9541403
Infect Immun. 2007 Jan;75(1):211-9
17074852
Nurs Times. 2005 May 17-23;101(20):42
15918462
Eukaryot Cell. 2003 Feb;2(1):84-94
12582125
Comput Appl Biosci. 1997 Aug;13(4):397-406
9283754
Toxicol Pathol. 1999 Nov-Dec;27(6):607-17
10588540