title: Aptamer beacons for visualization of endogenous protein HIV-1 reverse transcriptase in living cells.
abstract: Direct visualization of endogenous proteins in living cells remains a challenge. Aptamer beacon is a promising technique to resolve this problem by combining the excellent protein binding specificity of the aptamer with the sensitive signal transduction mechanism of the molecular beacon. In this study, aptamer 93 del against HIV-1 reverse transcriptase (RT) was engineered into aptamer beacons to recognize and image HIV-1 RT. The constructed aptamer beacons could specifically bind to HIV-1 RT and the beacon-RT binding showed effective fluorescence signal transduction in homogeneous solution. In solutions with 1 μM of the aptamer beacon, the effective fluorescence signal increased with increasing concentration of HIV-1 RT from 0.5 μM to 5 μM. When the aptamer beacons were delivered into the living cells that transiently expressed HIV-1 RT, HIV-1 RT could be specifically labeled and imaged. The designed aptamer beacons were further successfully applied for RT imaging in HIV-1 integrated U1 cells. The method developed here may be extended to visualize many other endogenous proteins in living cells using appropriate aptamer beacons.
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liang
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Zhang
authors.forename: Hongping
authors.initials: H
authors.lastname: Wei
authors.forename: Qinxue
authors.initials: Q
authors.lastname: Hu
authors.forename: Jiaoyu
authors.initials: J
authors.lastname: Deng
authors.forename: Deyin
authors.initials: D
authors.lastname: Guo
authors.forename: Zongqiang
authors.initials: Z
authors.lastname: Cui
authors.forename: Xian-En
authors.initials: XE
authors.lastname: Zhang
mesh_terms: D002470:Cell Survival; D005455:Fluorescent Antibody Technique; D054303:HIV Reverse Transcriptase / Q000032:analysis*; D006367:HeLa Cells; D006801:Humans; D015345:Oligonucleotide Probes / Q000378:metabolism; D011485:Protein Binding; D052156:SELEX Aptamer Technique / Q000379:methods*; D015398:Signal Transduction
chemical_list: D015345:Oligonucleotide Probes; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase