Gene knockdown by large circular antisense for high-throughput functional genomics

Yun-Han LeeIk-Jae MoonBin HurJeong-Hoh ParkKil-Hwan HanSeok-Yong UhmYong-Joo KimKoo-Jeong KangJong-Wook ParkYoung-Bae SeuYoung-Ho KimJong-Gu Park
Dept. of Surgery (외과학); Dept. of Immunology (면역학); Dept. of Molecular Medicine (분자의학)
Issue Date
Nat Biotechnol, Vol.23(5) : 591-599, 2005
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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1. Journal Papers (연구논문) > 1. School of Medicine (의과대학) > Dept. of Surgery (외과학)
1. Journal Papers (연구논문) > 1. School of Medicine (의과대학) > Dept. of Immunology (면역학)
1. Journal Papers (연구논문) > 1. School of Medicine (의과대학) > Dept. of Molecular Medicine (분자의학)
Keimyung Author(s)
강구정; 박종욱; 박종구
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