Transchip: single-molecule detection of transcriptional elongation complexes.
A new single-molecule system, Transchip, was developed for analysis of transcription products at their genomic origins. The bacteriophage T7 RNA polymerase and its promoters were used in a model system, and resultant RNAs were imaged and detected at their positions along single template DNA molecules. The Transchip system has drawn from critical aspects of Optical Mapping, a single-molecule system that enables the construction of high-resolution ordered restriction maps of whole genomes from single DNA molecules. Through statistical analysis of hundreds of single-molecule template/transcript complexes, Transchip enables analysis of the locations and strength of promoters, the direction and processivity of transcription reactions, and the termination of transcription. These novel results suggest that the new system may serve as a high-throughput platform to investigate transcriptional events on a large genome-wide scale.
Wu
Tian
T
Laboratory for Molecular and Computational Genomics, Department of Chemistry, Laboratory of Genetics, University of Wisconsin-Madison, Madison, WI 53706, USA.
Schwartz
David C
DC
D017123:Bacteriophage T7 / Q000235:genetics; D001583:Benzoxazoles; D003001:Cloning, Molecular; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005456:Fluorescent Dyes; D011401:Promoter Regions, Genetic; D011806:Quinolinium Compounds; D013698:Templates, Genetic; D014158:Transcription, Genetic; D035602:Transcriptional Elongation Factors / Q000032:analysis*; D014764:Viral Proteins / Q000378:metabolism
D001583:Benzoxazoles; D017931:DNA Primers; D005456:Fluorescent Dyes; D011806:Quinolinium Compounds; D035602:Transcriptional Elongation Factors; D014764:Viral Proteins; C075296:1,1'-((4,4,7,7-tetramethyl)-4,7-diazaundecamethylene)bis-4-(3-methyl-2,3-dihydro(benzo-1,3-oxazole)-2-methylidene)quinolinium; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases