Literature DB >> 22974138

Release of intracellular proteins by electroporation with preserved cell viability.

Yihong Zhan1, Chen Sun, Zhenning Cao, Ning Bao, Jianhua Xing, Chang Lu.   

Abstract

Extraction of intracellular proteins from cells is often an important first step for conducting molecular biology and proteomics studies. Although ultrasensitive detection and analytical technology at the single molecule level is becoming routine, protein extraction techniques have not followed suit and still call for complete lysis that leads to cell death. In principle, with refined extraction techniques, intracellular proteins can potentially be extracted without killing the cell. In this Letter, we demonstrate that electroporation is capable of releasing intracellular proteins from adherent Chinese hamster ovary cells while preserving the cell viability. By tuning the duration and intensity of an electric pulse, we were able to control the average amount of protein release and the percentage of viable cells after the operation. Our results indicate that a substantial fraction of the cell population was able to release proteins under electroporation and survive the procedure. Interestingly, at the single cell level, the probability for cell death does not increase with more protein release. This work paves the way to extracting and analyzing intracellular proteins while keeping cells live.

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Year:  2012        PMID: 22974138     DOI: 10.1021/ac302462s

Source DB:  PubMed          Journal:  Anal Chem        ISSN: 0003-2700            Impact factor:   6.986


  11 in total

1.  Protein Extraction by Means of Electroporation from E. coli with Preserved Viability.

Authors:  Sasa Haberl Meglic; Tilen Marolt; Damijan Miklavcic
Journal:  J Membr Biol       Date:  2015-07-23       Impact factor: 1.843

2.  A mechanical cell disruption microfluidic platform based on an on-chip micropump.

Authors:  Yinuo Cheng; Yue Wang; Zhiyuan Wang; Liang Huang; Mingzhao Bi; Wenxiao Xu; Wenhui Wang; Xiongying Ye
Journal:  Biomicrofluidics       Date:  2017-04-04       Impact factor: 2.800

3.  Molecular extraction in single live cells by sneaking in and out magnetic nanomaterials.

Authors:  Zhen Yang; Liangzi Deng; Yucheng Lan; Xiaoliu Zhang; Zhonghong Gao; Ching-Wu Chu; Dong Cai; Zhifeng Ren
Journal:  Proc Natl Acad Sci U S A       Date:  2014-07-16       Impact factor: 11.205

4.  USNCTAM perspectives on mechanics in medicine.

Authors:  Gang Bao; Yuri Bazilevs; Jae-Hyun Chung; Paolo Decuzzi; Horacio D Espinosa; Mauro Ferrari; Huajian Gao; Shaolie S Hossain; Thomas J R Hughes; Roger D Kamm; Wing Kam Liu; Alison Marsden; Bernhard Schrefler
Journal:  J R Soc Interface       Date:  2014-08-06       Impact factor: 4.118

5.  Calcein Release from Cells In Vitro via Reversible and Irreversible Electroporation.

Authors:  Violeta Rajeckaitė; Baltramiejus Jakštys; Arnas Rafanavičius; Martynas Maciulevičius; Milda Jakutavičiūtė; Saulius Šatkauskas
Journal:  J Membr Biol       Date:  2017-11-15       Impact factor: 1.843

6.  Combined Numerical and Experimental Investigation of Localized Electroporation-Based Cell Transfection and Sampling.

Authors:  Prithvijit Mukherjee; S Shiva P Nathamgari; John A Kessler; Horacio D Espinosa
Journal:  ACS Nano       Date:  2018-11-27       Impact factor: 15.881

Review 7.  Microfluidic electroporation for cellular analysis and delivery.

Authors:  Tao Geng; Chang Lu
Journal:  Lab Chip       Date:  2013-10-07       Impact factor: 6.799

Review 8.  Micro- and Nanoscale Technologies for Delivery into Adherent Cells.

Authors:  Wonmo Kang; Rebecca L McNaughton; Horacio D Espinosa
Journal:  Trends Biotechnol       Date:  2016-06-07       Impact factor: 19.536

9.  Electroporation-delivered fluorescent protein biosensors for probing molecular activities in cells without genetic encoding.

Authors:  Chen Sun; Mingxing Ouyang; Zhenning Cao; Sai Ma; Hamzeh Alqublan; Nammalwar Sriranganathan; Yingxiao Wang; Chang Lu
Journal:  Chem Commun (Camb)       Date:  2014-10-09       Impact factor: 6.222

Review 10.  Cytoskeletal Disruption after Electroporation and Its Significance to Pulsed Electric Field Therapies.

Authors:  Philip M Graybill; Rafael V Davalos
Journal:  Cancers (Basel)       Date:  2020-04-30       Impact factor: 6.639

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