Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles

Abstract Given the growing interest in molecular diagnosis, highly extensive and selective detection of genetic targets from a very limited amount of samples is in high demand. We demonstrated the highly sensitive and multiplexed one-step RT-qPCR platform for RNA analysis using microparticles as ind...

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Autores principales:	Mi Yeon Kim, Seungwon Jung, Junsun Kim, Heon Jeong Lee, Seunghwa Jeong, Sang Jun Sim, Sang Kyung Kim
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/02b5303dbf70414898ee8fc6b1350172
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spelling	oai:doaj.org-article:02b5303dbf70414898ee8fc6b13501722021-12-02T11:39:33ZHighly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles10.1038/s41598-021-85728-y2045-2322https://doaj.org/article/02b5303dbf70414898ee8fc6b13501722021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85728-yhttps://doaj.org/toc/2045-2322Abstract Given the growing interest in molecular diagnosis, highly extensive and selective detection of genetic targets from a very limited amount of samples is in high demand. We demonstrated the highly sensitive and multiplexed one-step RT-qPCR platform for RNA analysis using microparticles as individual reactors. Those particles are equipped with a controlled release system of thermo-responsive materials, and are able to capture RNA targets inside. The particle-based assay can successfully quantify multiple target RNAs from only 200 pg of total RNA. The assay can also quantify target RNAs from a single cell with the aid of a pre-concentration process. We carried out 8-plex one-step RT-qPCR using tens of microparticles, which allowed extensive mRNA profiling. The circadian cycles were shown by the multiplex one-step RT-qPCR in human cell and human hair follicles. Reliable 24-plex one-step RT-qPCR was developed using a single operation in a PCR chip without any loss of performance (i.e., selectivity and sensitivity), even from a single hair. Many other disease-related transcripts can be monitored using this versatile platform. It can also be used non–invasively for samples obtained in clinics.Mi Yeon KimSeungwon JungJunsun KimHeon Jeong LeeSeunghwa JeongSang Jun SimSang Kyung KimNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mi Yeon Kim Seungwon Jung Junsun Kim Heon Jeong Lee Seunghwa Jeong Sang Jun Sim Sang Kyung Kim Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles
description	Abstract Given the growing interest in molecular diagnosis, highly extensive and selective detection of genetic targets from a very limited amount of samples is in high demand. We demonstrated the highly sensitive and multiplexed one-step RT-qPCR platform for RNA analysis using microparticles as individual reactors. Those particles are equipped with a controlled release system of thermo-responsive materials, and are able to capture RNA targets inside. The particle-based assay can successfully quantify multiple target RNAs from only 200 pg of total RNA. The assay can also quantify target RNAs from a single cell with the aid of a pre-concentration process. We carried out 8-plex one-step RT-qPCR using tens of microparticles, which allowed extensive mRNA profiling. The circadian cycles were shown by the multiplex one-step RT-qPCR in human cell and human hair follicles. Reliable 24-plex one-step RT-qPCR was developed using a single operation in a PCR chip without any loss of performance (i.e., selectivity and sensitivity), even from a single hair. Many other disease-related transcripts can be monitored using this versatile platform. It can also be used non–invasively for samples obtained in clinics.
format	article
author	Mi Yeon Kim Seungwon Jung Junsun Kim Heon Jeong Lee Seunghwa Jeong Sang Jun Sim Sang Kyung Kim
author_facet	Mi Yeon Kim Seungwon Jung Junsun Kim Heon Jeong Lee Seunghwa Jeong Sang Jun Sim Sang Kyung Kim
author_sort	Mi Yeon Kim
title	Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles
title_short	Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles
title_full	Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles
title_fullStr	Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles
title_full_unstemmed	Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles
title_sort	highly sensitive and multiplexed one-step rt-qpcr for profiling genes involved in the circadian rhythm using microparticles
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/02b5303dbf70414898ee8fc6b1350172
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Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles

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