Abstract：
:We developed genetically encoded RNA probes for characterizing localization and dynamics of mitochondrial RNA (mtRNA) in single living cells. The probes consist of two RNA-binding domains of PUMILIO1, each connected with split fragments of a fluorescent protein capable of reconstituting upon binding to a target RNA. We designed the probes to specifically recognize a 16-base sequence of mtRNA encoding NADH dehydrogenase subunit 6 (ND6) and to be targeted into the mitochondrial matrix, which allowed real-time imaging of ND6 mtRNA localization in living cells. We showed that ND6 mtRNA is localized within mitochondria and concentrated particularly on mitochondrial DNA (mtDNA). Movement of the ND6 mtRNA is restricted but oxidative stress induces the mtRNA to disperse in the mitochondria and gradually decompose. These probes provide a means to study spatial and temporal mRNA dynamics in intracellular compartments in living mammalian cells.
journal_name
Nat Methodsjournal_title
Nature methodsauthors
Ozawa T,Natori Y,Sato M,Umezawa Ydoi
10.1038/nmeth1030subject
Has Abstractpub_date
2007-05-01 00:00:00pages
413-9issue
5eissn
1548-7091issn
1548-7105pii
nmeth1030journal_volume
4pub_type
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