Electroporation and Microinjection Successfully Deliver Single-Stranded and Duplex DNA into Live Cells as Detected by FRET Measurements

被引：12

作者：

Bamford, Rosemary A. ^{[1
]}

Zhao, Zheng-yun ^{[2
]}

Hotchin, Neil A. ^{[3
]}

Styles, Iain B. ^{[4
]}

Nash, Gerard B. ^{[5
]}

Tucker, James H. R. ^{[2
]}

Bicknell, Roy ^{[6
]}

机构：

[1] Univ Birmingham, PSIBS Doctoral Training Ctr, Birmingham B15 2TT, W Midlands, England

[2] Univ Birmingham, Sch Chem, Birmingham B15 2TT, W Midlands, England

[3] Univ Birmingham, Sch Biosci, Birmingham B15 2TT, W Midlands, England

[4] Univ Birmingham, Sch Comp Sci, Birmingham B15 2TT, W Midlands, England

[5] Univ Birmingham, Sch Clin & Expt Med, Ctr Cardiovasc Sci, Birmingham B15 2TT, W Midlands, England

[6] Univ Birmingham, Inst Biomed Res, Sch Immun & Infect, Birmingham B15 2TT, W Midlands, England

来源：

PLOS ONE | 2014年 / 9卷 / 04期

基金：

英国工程与自然科学研究理事会;

关键词：

RESONANCE ENERGY-TRANSFER; NUCLEIC-ACID HYBRIDIZATION; MOLECULAR BEACONS; LIVING CELLS; INTRACELLULAR DELIVERY; MESSENGER-RNA; IN-VIVO; FLUORESCENCE; PROBES; ANTISENSE;

D O I：

10.1371/journal.pone.0095097

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Forster resonance energy transfer (FRET) technology relies on the close proximity of two compatible fluorophores for energy transfer. Tagged (Cy3 and Cy5) complementary DNA strands forming a stable duplex and a doubly-tagged single strand were shown to demonstrate FRET outside of a cellular environment. FRET was also observed after transfecting these DNA strands into fixed and live cells using methods such as microinjection and electroporation, but not when using lipid based transfection reagents, unless in the presence of the endosomal acidification inhibitor bafilomycin. Avoiding the endocytosis pathway is essential for efficient delivery of intact DNA probes into cells.

引用

页数：8

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