3C Technologies in plants.
Chromosome conformation capture (3C) and 3C-based technology have revolutionized studies on chromosomal interactions and their role in gene regulation and chromosome organization. 3C allows the in vivo identification of physical interactions between chromosomal regions. Such interactions are shown to play a role in various aspects of gene regulation, for example transcriptional activation of genes by remote enhancer sequences, or the silencing by Polycomb-group complexes. The last few years the number of publications involving chromosomal interactions increased significantly. Until now, however, the vast majority of the studies reported are performed in yeast or animal systems. So far, studies on plant systems are extremely limited, possibly due to the plant-specific characteristics that hamper the implementation of the 3C technique. In this paper we provide a plant-specific 3C protocol, optimized for maize tissue, and an extensive discussion on (i) plant-specific adjustments to the protocol, and (ii) solutions to problems that may arise when optimizing the protocol for the tissue or plant of interest. Together, this paper should facilitate the application of 3C technology to plant tissue and stimulate studies on the 3D conformation of chromosomal regions and chromosomes in plants.
Swammerdam Institute for Life Sciences, Universiteit van Amsterdam, Amsterdam, The Netherlands.
Iris
I
Hövel
Marieke
M
Louwers
Maike
M
Stam
D002467:Cell Nucleus / Q000235:genetics; D053144:Cell Nucleus Shape; D002874:Chromosome Mapping / Q000379:methods* / Q000592:standards; D032461:Chromosomes, Plant / Q000235:genetics*; D003432:Cross-Linking Reagents / Q000737:chemistry; D053837:DNA Cleavage; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D018745:Genome, Plant; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D012015:Reference Standards; D016707:Tissue Fixation / Q000379:methods; D003313:Zea mays / Q000166:cytology / Q000235:genetics*
D003432:Cross-Linking Reagents; D018744:DNA, Plant; D005404:Fixatives; D005557:Formaldehyde