Abstract
Gene silencing has been used widely in gene function studies and crop plant modification. Long hairpin RNA (lhRNA) results in high efficiency of gene silencing; however, constructing multiple lhRNA vectors using traditional approaches is both time consuming and costly. Also, most of the existing approaches are based on sequence-specific cloning of individual sequences and are therefore not suitable for preparing hpRNA libraries from a pool of mixed target sequences. The rolling circle amplification (RCA)-mediated hairpin RNA (RMHR) construction system is suitable for generating hairpin libraries from any gene of interest or pool of genes. Using the RMHR system, a long-hairpin RNA (lhRNA) library is generated from an Arabidopsis cDNA population containing known and unknown genes. Our results indicate that the RMHR technique permits the rapid, efficient, and low-cost preparation of genome-wide lhRNA expression libraries.
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Acknowledgments
This work was supported by National Natural Science Foundation of China (31000576) and the National Key Basic Research Program (2010CB125903) and Genetically Modified Organisms Major Breeding Projects (2009ZX08009-016B and 2010ZX08010-002).
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Wang, L., Fan, YL. (2012). Rolling Circle Amplification-Mediated Long Hairpin RNA Library Construction in Plants. In: Watson, J., Wang, MB. (eds) Antiviral Resistance in Plants. Methods in Molecular Biology, vol 894. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-882-5_21
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DOI: https://doi.org/10.1007/978-1-61779-882-5_21
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