Simultaneous imaging of microRNA or mRNA territories with protein territory in mammalian cells at single cell resolution

Ranjan, Amaresh K.; Joglekar, Mugdha V.; Atre, Ashwini; Patole, Milind S.; Bhonde, Ramesh; Hardikar, Anandwardhan A.

doi:10.4161/rna.20484

Cited by 10 publications

(11 citation statements)

References 23 publications

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“…The biogenesis of mitochondria involves replication of mitochondrial DNA and increase in their copy number. To study biogenesis in fixed tissue samples we devised a technique of "in situ mitochondrial DNA PCR" by modifying our previously developed protocol of in situ PCR for detection of transcripts and miRNA 51 . We carried out in situ PCR using a mitochondrial gene; MT-ATP8 specific TaqMan probe and primers in day 7 post MCAO tissue samples.…”

Section: Discussionmentioning

confidence: 99%

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Sovateltide (IRL-1620) activates neuronal differentiation and prevents mitochondrial dysfunction in adult mammalian brains following stroke

Ranjan

Briyal

Gulati

2020

Sci Rep

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The development of effective drugs for stroke is urgently required as it is the 2nd largest killer in the world and its incidence is likely to increase in the future. We have demonstrated cerebral endothelin B receptors (ETBR) as a potential target to treat acute cerebral ischemic stroke. However, the mechanism of ETBR mediated neural regeneration and repair remains elusive. In this study, a permanent middle cerebral artery occluded (MCAO) rat model was used. Sovateltide (an ETBR agonist) injected intravenously showed better survival and neurological and motor function improvement than control. Higher neuronal progenitor cells (NPCs) differentiation along with better mitochondrial morphology and biogenesis in the brain of sovateltide rats were noted. Exposure of cultured NPCs to hypoxia and sovateltide also showed higher NPC differentiation and maturation. This study shows a novel role of ETBR in NPCs and mitochondrial fate determination in cerebral ischemia, and in improving neurological deficit after stroke.

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Section: Discussionmentioning

confidence: 99%

“…In situ PCR was carried out to assess the mitochondrial biogenesis in the rat brain tissues, with following our previously developed in situ PCR protocol aimed to detect transcripts and miRNA in cultured cells 51 . In brief, the right brain hemisphere was dissected as depicted in Fig.…”

Section: Methodsmentioning

confidence: 99%

Sovateltide (IRL-1620) activates neuronal differentiation and prevents mitochondrial dysfunction in adult mammalian brains following stroke

Ranjan

Briyal

Gulati

2020

Sci Rep

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“…Although a number of new methods have been developed for determining miRNAs in single cells, they also fail to provide subcellular resolution. 29 – 33 …”

Section: Introductionmentioning

confidence: 99%

Probing cytoplasmic and nuclear microRNAs in single living cells via plasmonic affinity sandwich assay

Wen

Chen

et al. 2018

Chem. Sci.

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“…Manuscript to be reviewed combining in-situ TaqMan PCR with immunostaining to visualize protein expression (Ranjan et al 2012). This study however, relied on in-situ PCR and the availability of antibodies.…”

Section: Discussionmentioning

confidence: 99%

Peer Review #1 of "qPCR multiplex detection of microRNA and messenger RNA in a single reaction (v0.1)"

2020

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Reverse Transcription-Quantitative PCR (RT-qPCR) is one of the standards for analytical measurement of different RNA species in biological models. However, current Reverse Transcription (RT) based priming strategies are unable to synthesize differing RNAs and ncRNAs especially miRNAs, within a single tube. We present a new methodology, referred to as RNAmp, that measures in parallel miRNA and mRNA expression. We demonstrate this in various cell lines, then evaluate clinical utility by quantifying several miRNAs and mRNA simultaneously in sera. PCR efficiency in RNAmp was estimated between 1.8 and 1.9 which is comparable to standard miRNA and random primer RT approaches. Furthermore, when using RNAmp to detect selected mRNA and miRNAs, the quantification cycle (Cq) was several cycles lower. This low volume single-tube duplex protocol reduces technical variation and reagent usage and is suitable for uniform analysis of single or multiple miRNAs and/or mRNAs within a single qPCR reaction.

show abstract

Simultaneous imaging of microRNA or mRNA territories with protein territory in mammalian cells at single cell resolution

Cited by 10 publications

References 23 publications

Sovateltide (IRL-1620) activates neuronal differentiation and prevents mitochondrial dysfunction in adult mammalian brains following stroke

Sovateltide (IRL-1620) activates neuronal differentiation and prevents mitochondrial dysfunction in adult mammalian brains following stroke

Probing cytoplasmic and nuclear microRNAs in single living cells via plasmonic affinity sandwich assay

Peer Review #1 of "qPCR multiplex detection of microRNA and messenger RNA in a single reaction (v0.1)"

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