Methods to Investigate the Regulatory Role of Small RNAs and Ribosomal Occupancy of Plasmodium falciparum.
Journal Article
The genetic variation responsible for the sickle cell allele (HbS) enables erythrocytes to resist infection by the malaria parasite, P. falciparum. The molecular basis of this resistance, which is known to be multifactorial, remains incompletely understood. Recent studies found that the differential expression of erythrocyte microRNAs, once translocated into malaria parasites, affect both gene regulation and parasite growth. These miRNAs were later shown to inhibit mRNA translation by forming a chimeric RNA transcript via 5' RNA fusion with discreet subsets of parasite mRNAs. Here, the techniques that were used to study the functional role and putative mechanism underlying erythrocyte microRNAs on the gene regulation and translational potential of P. falciparum, including the transfection of modified synthetic microRNAs into host erythrocytes, will be detailed. Finally, a polysome gradient method is used to determine the extent of translation of these transcripts. Together, these techniques allowed us to demonstrate that the dysregulated levels of erythrocyte microRNAs contribute to cell-intrinsic malaria resistance of sickle erythrocytes.
Full Text
Duke Authors
Cited Authors
- LaMonte, G; Walzer, KA; Lacsina, J; Nicchitta, C; Chi, J-T
Published Date
- December 4, 2015
Published In
Start / End Page
- e53214 -
PubMed ID
- 26709459
Pubmed Central ID
- 26709459
Electronic International Standard Serial Number (EISSN)
- 1940-087X
Digital Object Identifier (DOI)
- 10.3791/53214
Language
- eng
Conference Location
- United States