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Gene knockdown by large circular antisense for high-throughput functional genomics.
- Source :
-
Nature biotechnology [Nat Biotechnol] 2005 May; Vol. 23 (5), pp. 591-9. Date of Electronic Publication: 2005 May 01. - Publication Year :
- 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-alpha, 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.
- Subjects :
- Animals
Bacteriophage M13 genetics
Cell Line, Tumor
Liver Neoplasms genetics
Mice
Neoplasm Proteins genetics
Chromosome Mapping methods
DNA, Antisense genetics
Gene Expression Profiling methods
Gene Silencing
Gene Targeting methods
Genomics methods
Liver Neoplasms metabolism
Neoplasm Proteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1087-0156
- Volume :
- 23
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Nature biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 15867911
- Full Text :
- https://doi.org/10.1038/nbt1089