In vivo cloning of artificial DNA nanostructures

被引:92
作者
Lin, Chenxiang [2 ,3 ]
Rinker, Sherri [2 ,3 ]
Wang, Xing [1 ]
Liu, Yan [2 ,3 ]
Seeman, Nadrian C. [1 ]
Yan, Hao [2 ,3 ]
机构
[1] NYU, Dept Chem, New York, NY 10003 USA
[2] Arizona State Univ, Dept Chem & Biochem, Tempe, AZ 85287 USA
[3] Arizona State Univ, Biodesign Inst, Tempe, AZ 85287 USA
关键词
DNA nanotechnology; immobile DNA junction; self-replication; synthetic biology;
D O I
10.1073/pnas.0805416105
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
Mimicking nature is both a key goal and a difficult challenge for the scientific enterprise. DNA, well known as the genetic-information carrier in nature, can be replicated efficiently in living cells. Today, despite the dramatic evolution of DNA nanotechnology, a versatile method that replicates artificial DNA nanostructures with complex secondary structures remains an appealing target. Previous success in replicating DNA nanostructures enzymatically in vitro suggests that a possible solution could be cloning these nanostructures by using viruses. Here, we report a system where a single-stranded DNA nanostructure (Holliday junction or paranemic cross-over DNA), is inserted into a phagemid, transformed into XL1-Blue cells and amplified in vivo in the presence of helper phages. High copy numbers of cloned nanostructures can be obtained readily by using standard molecular biology techniques. Correct replication is verified by a number of assays including nondenaturing PAGE, Ferguson analysis, endonuclease VII digestion, and hydroxyl radical autofootprinting. The simplicity, efficiency, and fidelity of nature are fully reflected in this system. UV-induced psoralen cross-linking is used to probe the secondary structure of the inserted junction in infected cells. Our data suggest the possible formation of the immobile four-arm junction in vivo.
引用
收藏
页码:17626 / 17631
页数:6
相关论文
共 44 条
[1]   Hydroxyl radical footprinting in vivo:: mapping macromolecular structures with synchrotron radiation [J].
Adilakshmi, Tadepalli ;
Lease, Richard A. ;
Woodson, Sarah A. .
NUCLEIC ACIDS RESEARCH, 2006, 34 (08)
[2]  
[Anonymous], 2001, Anal Biochem
[3]   SYNTHESIS FROM DNA OF A MOLECULE WITH THE CONNECTIVITY OF A CUBE [J].
CHEN, JH ;
SEEMAN, NC .
NATURE, 1991, 350 (6319) :631-633
[4]   CONSTRUCTION AND ANALYSIS OF MONOMOBILE DNA JUNCTIONS [J].
CHEN, JH ;
CHURCHILL, MEA ;
TULLIUS, TD ;
KALLENBACH, NR ;
SEEMAN, NC .
BIOCHEMISTRY, 1988, 27 (16) :6032-6038
[5]   DNA-directed assembly of single-wall carbon nanotubes [J].
Chen, Yi ;
Liu, Haipeng ;
Ye, Tao ;
Kim, Junghwa ;
Mao, Chengde .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2007, 129 (28) :8696-+
[6]   A HOLLIDAY RECOMBINATION INTERMEDIATE IS TWOFOLD SYMMETRIC [J].
CHURCHILL, MEA ;
TULLIUS, TD ;
KALLENBACH, NR ;
SEEMAN, NC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (13) :4653-4656
[7]   PSORALENS AS PHOTOACTIVE PROBES OF NUCLEIC-ACID STRUCTURE AND FUNCTION - ORGANIC-CHEMISTRY, PHOTOCHEMISTRY, AND BIOCHEMISTRY [J].
CIMINO, GD ;
GAMPER, HB ;
ISAACS, ST ;
HEARST, JE .
ANNUAL REVIEW OF BIOCHEMISTRY, 1985, 54 :1151-1193
[8]   DNA-nanotube-induced alignment of membrane proteins for NMR structure determination [J].
Douglas, Shawn M. ;
Chou, James J. ;
Shih, William M. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (16) :6644-6648
[9]   DNA nanotechnology: Chemical copying of connectivity [J].
Eckardt, LH ;
Naumann, K ;
Pankau, WM ;
Rein, M ;
Schweitzer, M ;
Windhab, N ;
von Kiedrowski, G .
NATURE, 2002, 420 (6913) :286-286
[10]  
ESPOSITO F, 1988, J BIOL CHEM, V263, P11466