Acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound

Abstract Efficient intracellular delivery of biologically active macromolecules has been a challenging but important process for manipulating live cells for research and therapeutic purposes. There have been limited transfection techniques that can deliver multiple types of active molecules simultan...

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Autores principales:	Sangpil Yoon, Pengzhi Wang, Qin Peng, Yingxiao Wang, K. Kirk Shung
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/be26cc80862341fdae4ced22fda1dc16
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spelling	oai:doaj.org-article:be26cc80862341fdae4ced22fda1dc162021-12-02T15:04:58ZAcoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound10.1038/s41598-017-05722-12045-2322https://doaj.org/article/be26cc80862341fdae4ced22fda1dc162017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05722-1https://doaj.org/toc/2045-2322Abstract Efficient intracellular delivery of biologically active macromolecules has been a challenging but important process for manipulating live cells for research and therapeutic purposes. There have been limited transfection techniques that can deliver multiple types of active molecules simultaneously into single-cells as well as different types of molecules into physically connected individual neighboring cells separately with high precision and low cytotoxicity. Here, a high frequency ultrasound-based remote intracellular delivery technique capable of delivery of multiple DNA plasmids, messenger RNAs, and recombinant proteins is developed to allow high spatiotemporal visualization and analysis of gene and protein expressions as well as single-cell gene editing using clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease (Cas9), a method called acoustic-transfection. Acoustic-transfection has advantages over typical sonoporation because acoustic-transfection utilizing ultra-high frequency ultrasound over 150 MHz can directly deliver gene and proteins into cytoplasm without microbubbles, which enables controlled and local intracellular delivery to acoustic-transfection technique. Acoustic-transfection was further demonstrated to deliver CRISPR-Cas9 systems to successfully modify and reprogram the genome of single live cells, providing the evidence of the acoustic-transfection technique for precise genome editing using CRISPR-Cas9.Sangpil YoonPengzhi WangQin PengYingxiao WangK. Kirk ShungNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sangpil Yoon Pengzhi Wang Qin Peng Yingxiao Wang K. Kirk Shung Acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound
description	Abstract Efficient intracellular delivery of biologically active macromolecules has been a challenging but important process for manipulating live cells for research and therapeutic purposes. There have been limited transfection techniques that can deliver multiple types of active molecules simultaneously into single-cells as well as different types of molecules into physically connected individual neighboring cells separately with high precision and low cytotoxicity. Here, a high frequency ultrasound-based remote intracellular delivery technique capable of delivery of multiple DNA plasmids, messenger RNAs, and recombinant proteins is developed to allow high spatiotemporal visualization and analysis of gene and protein expressions as well as single-cell gene editing using clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease (Cas9), a method called acoustic-transfection. Acoustic-transfection has advantages over typical sonoporation because acoustic-transfection utilizing ultra-high frequency ultrasound over 150 MHz can directly deliver gene and proteins into cytoplasm without microbubbles, which enables controlled and local intracellular delivery to acoustic-transfection technique. Acoustic-transfection was further demonstrated to deliver CRISPR-Cas9 systems to successfully modify and reprogram the genome of single live cells, providing the evidence of the acoustic-transfection technique for precise genome editing using CRISPR-Cas9.
format	article
author	Sangpil Yoon Pengzhi Wang Qin Peng Yingxiao Wang K. Kirk Shung
author_facet	Sangpil Yoon Pengzhi Wang Qin Peng Yingxiao Wang K. Kirk Shung
author_sort	Sangpil Yoon
title	Acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound
title_short	Acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound
title_full	Acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound
title_fullStr	Acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound
title_full_unstemmed	Acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound
title_sort	acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/be26cc80862341fdae4ced22fda1dc16
work_keys_str_mv	AT sangpilyoon acoustictransfectionforgenomicmanipulationofsinglecellsusinghighfrequencyultrasound AT pengzhiwang acoustictransfectionforgenomicmanipulationofsinglecellsusinghighfrequencyultrasound AT qinpeng acoustictransfectionforgenomicmanipulationofsinglecellsusinghighfrequencyultrasound AT yingxiaowang acoustictransfectionforgenomicmanipulationofsinglecellsusinghighfrequencyultrasound AT kkirkshung acoustictransfectionforgenomicmanipulationofsinglecellsusinghighfrequencyultrasound
_version_	1718388980053966848

Acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound

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