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Rolling Circle Amplification-Mediated Long Hairpin RNA Library Construction in Plants

 Last Update: 2020-11-20
 Source: Internet
 Author: User

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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
c
DNA
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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