Native chromatin immunoprecipitation (N-ChIP) and ChIP-Seq of Schistosoma mansoni: Critical experimental parameters.
Histone modifications are important epigenetic marks that influence chromatin structure and consequently play a role in the control of eukaryotic transcription. Several histone modifying enzymes have been characterized in Schistosoma mansoni and it has been suggested that the regulation of gene transcription in schistosomes may require the action of these enzymes. However, the influence of chromatin structure on gene transcription in schistosomes has never been investigated. Chromatin immunoprecipitation (ChIP) is the technique of choice to study the relationship between histone modifications and gene expression. Although this technique has been widely used with cultured cells from model organisms and with many unicellular organisms, it remains challenging to apply this technique to non-conventional organisms that undergo complex life cycles. In this work, we describe a native ChIP procedure that is applicable to all the stages of the S. mansoni life cycle and does not require expensive equipment. Immunoprecipitated DNA was analysed on a whole-genome scale using massively parallel sequencing (ChIP-Sequencing or ChIP-Seq). We show that ChIP-Seq and conventional quantitative PCR deliver comparable results for a life-cycle regulated locus, smRHO, that encodes a guanine-protein coupled receptor. This is the first time that the ChIP-Seq procedure has been applied to a parasite. This technique opens new ways for analyzing epigenetic mechanisms in S. mansoni at a whole-genome scale and on the level of individual loci.
Cosseau
Céline
C
Parasitologie Fonctionnelle et Evolutive, UMR 5244, CNRS EPHE Université de Perpignan, Perpignan Cedex, France.
Azzi
Abdelhalim
A
Smith
Kristina
K
Freitag
Michael
M
Mitta
Guillaume
G
Grunau
Christoph
C
D000818:Animals; D000909:Antibodies, Helminth / Q000378:metabolism; D002478:Cells, Cultured; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D006657:Histones / Q000378:metabolism; D006801:Humans; D008018:Life Cycle Stages; D011485:Protein Binding; D015203:Reproducibility of Results; D012243:Rhodopsin / Q000235:genetics; D012550:Schistosoma mansoni / Q000235:genetics* / Q000378:metabolism
D000909:Antibodies, Helminth; D006657:Histones; D012243:Rhodopsin