Membrane-Embedded Channel of Bacteriophage Phi29 DNA-Packaging Motor for Translocation and Sensing of Double-Stranded DNA - Nanopores: Sensing and Fundamental Biological Interactions - page 103

4.7.3 Nanopore Based Sequencing of DNA
Nanopore-based DNA sequencing applications holds considerable promise and has
the potential to operate orders of magnitude faster and cheaper than traditional
methods. In addition, this is a label-free, single molecule recognition approach
and requires no sample preparation or amplification. To date, there have been a
number of reports demonstrating successful applications of nanopore-based nucleic
acid analysis using either protein pores or solid state analogs [ 23 - 26 , 28 - 31 , 56 ].
However, the limiting factor for the single pore DNA sequencing technology is
that the ultra-fast DNA passage is beyond the temporal resolution of currently
available optical and electrical technologies for detecting individual nucleotides
with high sensitivity and confidence. The key challenge therefore is to be able
to slow the passage of DNA in a controlled fashion while maintaining high signal-
to-noise ratio for accurately discriminating the bases. The large channel of the
phi29 connector provides room for modifications, conjugations and placement of
barriers for better signal detection.
4.8 Conclusions
Herein, an engineered form of the phi29 connector has been incorporated into a
lipid bilayer, forming a highly conductive nanopore. In addition, the connector is
a biological pore that is extremely reproducible and easily engineered, making
it ideal and suitable for future biomedical applications. The robust properties of
the phi29 connector with a large uniform size support the expectation that viral
components are a new generation of nanomaterials or nanoscale building blocks.
Acknowledgments We thank Feng Xiao and Ying Cai for constructing the recombinant
connectors; Rong Zhang for Q-PCR analysis, Jing Peng and Jia Geng for assistance in BLM
experiments, and Dan Shu for the in vitro packaging assays. The research was supported by NIH
grants GM059944, EB003730, and NIH Nanomedicine Development Center: Phi29 DNA
Packaging Motor for Nanomedicine, through the NIH Roadmap for Medical Research (PN2 EY
018230) to P.G., who is also a co-founder of Kylin Therapeutics, Inc.
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