Gene knockdown by large circular antisense for high-throughput functional genomics
- Affiliated Author(s)
- 강구정; 박종욱; 박종구
- Alternative Author(s)
- Kang, Koo Jeong; Park, Jong Wook; Park, Jong Gu
- Journal Title
- Nat Biotechnol
- ISSN
- 1087-0156
- Issued Date
- 2005
- Abstract
- Single-stranded genomic DNA of recombinant M13 phages was tested as an antisense molecule and examined for its
usefulness in high-throughput functional genomics. cDNA fragments of various genes (TNF-a, c-myc, c-myb, cdk2 and cdk4)
were independently cloned into phagemid vectors. Using the life cycle of M13 bacteriophages, large circular (LC)-molecules,
antisense to their respective genes, were prepared from the culture supernatant of bacterial transformants. LC-antisense
molecules exhibited enhanced stability, target specificity and no need for target-site searches. High-throughput functional
genomics was then attempted with an LC-antisense library, which was generated by using a phagemid vector that incorporated
a unidirectional subtracted cDNA library derived from liver cancer tissue. We identified 56 genes involved in the growth of
these cells. These results indicate that an antisense sequence as a part of single-stranded LC-genomic DNA of recombinant
M13 phages exhibits effective antisense activity, and may have potential for high-throughput functional genomics.
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