Fluorescent in situ hybridization of mitochondrial DNA and RNA.
To reveal nucleic acid localization in mitochondria, we designed molecular beacon fluorescent probes against: i) the light strand complementary to ND5 mitochondrial DNA (mtDNA) gene (annealing also to corresponding mRNA); ii) displacement (D) loop 7S DNA (annealing also to parallel heavy strand mtDNA and corresponding light strand transcript); iii) the proximal D-loop heavy strand displaced by the light strand promoter minor RNA. Confocal microscopy demonstrated ND5 probe spreading (less for other probes) in mitochondrial reticulum tubules but upon RNase A treatment all probes contoured mtDNA nucleoid localization. DNase I spread the signal over mitochondrial tubules. Future applications are discussed.
Department No. 75, Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
Lukáš
L
Alán
Jaroslav
J
Zelenka
Jan
J
Ježek
Andrea
A
Dlasková
Petr
P
Ježek
D045744:Cell Line, Tumor; D004272:DNA, Mitochondrial / Q000378:metabolism*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D018613:Microscopy, Confocal; D012313:RNA / Q000378:metabolism*; D000077278:RNA, Mitochondrial
D004272:DNA, Mitochondrial; D000077278:RNA, Mitochondrial; D012313:RNA