Translocating Kilobase RNA through the Staphylococcal α-Hemolysin Nanopore

被引：49

作者：

Cracknell, James A. ^{[1
]}

Japrung, Deanpen ^{[1
]}

Bayley, Hagan ^{[1
]}

机构：

[1] Univ Oxford, Dept Chem, Chem Res Lab, Oxford OX1 3TA, England

来源：

NANO LETTERS | 2013年 / 13卷 / 06期

基金：

美国国家卫生研究院;

关键词：

alpha-Hemolysin; RNA; translocation; nanopore; heteropolymer; single-molecule; SINGLE-NUCLEOTIDE RESOLUTION; NUCLEIC-ACID MOLECULES; STRANDED-DNA; STATE; DISCRIMINATION; UREA; OLIGONUCLEOTIDES; TRANSPORT; SECONDARY; SENSORS;

D O I：

10.1021/nl400560r

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

The electrophoretic translocation of polynucleotides through nanopores may permit direct single-molecule nucleic acid sequencing. Here we describe the translocation of ssRNA heteropolymers (91-6083 bases) through the alpha-hemolysin nanopore. Translocation of these long ssRNAs is characterized by surprisingly long, almost complete ionic current blockades with durations averaging milliseconds per base (at +180 mV). The event durations decrease exponentially with increased transmembrane potential but are largely unaffected by the presence of urea. When the ssRNA is coupled at the 3' end to streptavidin, which cannot translocate through the pore, permanent blockades are observed, supporting our conclusion that the transient blockades arise from ssRNA translocation.

引用

页码：2500 / 2505

页数：6

共 45 条

[1] Microsecond time-scale discrimination among polycytidylic acid, polyadenylic acid, and polyuridylic acid as homopolymers or as segments within single RNA molecules [J].

Akeson, M ;

Branton, D ;

Kasianowicz, JJ ;

Brandin, E ;

Deamer, DW .

BIOPHYSICAL JOURNAL, 1999, 77 (06) :3227-3233

[2] Individual RNA Base Recognition in Immobilized Oligonucleotides Using a Protein Nanopore [J].

Ayub, Mariam ;

Bayley, Hagan .

NANO LETTERS, 2012, 12 (11) :5637-5643

[3] Stochastic sensors inspired by biology [J].

Bayley, H ;

Cremer, PS .

NATURE, 2001, 413 (6852) :226-230

[4] Are we there yet? Comment on "Nanopores: A journey towards DNA sequencing" by Meni Wanunu [J].

Bayley, Hagan .

PHYSICS OF LIFE REVIEWS, 2012, 9 (02) :161-163

[5] The potential and challenges of nanopore sequencing [J].

Branton, Daniel ;

Deamer, David W. ;

Marziali, Andre ;

Bayley, Hagan ;

Benner, Steven A. ;

Butler, Thomas ;

Di Ventra, Massimiliano ;

Garaj, Slaven ;

Hibbs, Andrew ;

Huang, Xiaohua ;

Jovanovich, Stevan B. ;

Krstic, Predrag S. ;

Lindsay, Stuart ;

Ling, Xinsheng Sean ;

Mastrangelo, Carlos H. ;

Meller, Amit ;

Oliver, John S. ;

Pershin, Yuriy V. ;

Ramsey, J. Michael ;

Riehn, Robert ;

Soni, Gautam V. ;

Tabard-Cossa, Vincent ;

Wanunu, Meni ;

Wiggin, Matthew ;

Schloss, Jeffery A. .

NATURE BIOTECHNOLOGY, 2008, 26 (10) :1146-1153

[6] Ionic current blockades from DNA and RNA molecules in the α-hemolysin nanopore [J].

Butler, Tom Z. ;

Gundlach, Jens H. ;

Troll, Mark .

BIOPHYSICAL JOURNAL, 2007, 93 (09) :3229-3240

[7] Single-molecule DNA detection with an engineered MspA protein nanopore [J].

Butler, Tom Z. ;

Pavlenok, Mikhail ;

Derrington, Ian M. ;

Niederweis, Michael ;

Gundlach, Jens H. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (52) :20647-20652

[8] Confinement-driven translocation of a flexible polymer [J].

Cacciuto, Angelo ;

Luijten, Erik .

PHYSICAL REVIEW LETTERS, 2006, 96 (23)

[9] Spontaneous oligomerization of a staphylococcal α-hemolysin conformationally constrained by removal of residues that form the transmembrane β-barrel [J].

Cheley, S ;

Malghani, MS ;

Song, LZ ;

Hobaugh, M ;

Gouaux, JE ;

Yang, J ;

Bayley, H .

PROTEIN ENGINEERING, 1997, 10 (12) :1433-1443

[10] Automated forward and reverse ratcheting of DNA in a nanopore at 5-Å precision [J].

Cherf, Gerald M. ;

Lieberman, Kate R. ;

Rashid, Hytham ;

Lam, Christopher E. ;

Karplus, Kevin ;

Akeson, Mark .

NATURE BIOTECHNOLOGY, 2012, 30 (04) :344-348

← 1 2 3 4 5 →