2013
DOI: 10.1007/s10529-013-1197-3
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Universal vectors for constructing artificial microRNAs in plants

Abstract: Universal amiRNA vectors (pUAs) for constructing plant amiRNAs in Arabidopsis and rice have been developed. By using type IIg restriction enzyme, BaeI, a single amiRNA construct can be produced using only one PCR and one ligation reaction. Thus, only one pair of primers is required for each amiRNA vector and these can be designed to be compatible with existing or newly developed methods. Because the BaeI recognition sequence is completely digested, there is no modification to the miRNA backbone, therefore avoi… Show more

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Cited by 5 publications
(3 citation statements)
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“…Nonlinearized plasmid molecules with no amiRNA insert fail to propagate in E. coli ccdB-sensitive strains, such as DH5a or DH10B. In summary, compared with other amiRNA cloning methods (Schwab et al, 2006;Qu et al, 2007;Chen et al, 2009;Molnar et al, 2009;Wang et al, 2010Wang et al, , 2012Eamens et al, 2011;Yan et al, 2011;Liang et al, 2012;Zhou et al, 2013), this method is relatively simple, fast, and cost effective (Fig. 2C).…”
Section: Atmir390a-based Vectorsmentioning
confidence: 92%
See 1 more Smart Citation
“…Nonlinearized plasmid molecules with no amiRNA insert fail to propagate in E. coli ccdB-sensitive strains, such as DH5a or DH10B. In summary, compared with other amiRNA cloning methods (Schwab et al, 2006;Qu et al, 2007;Chen et al, 2009;Molnar et al, 2009;Wang et al, 2010Wang et al, , 2012Eamens et al, 2011;Yan et al, 2011;Liang et al, 2012;Zhou et al, 2013), this method is relatively simple, fast, and cost effective (Fig. 2C).…”
Section: Atmir390a-based Vectorsmentioning
confidence: 92%
“…This prevents vector self-ligation and eliminates the need to modify the ends of insert oligonucleotide sequences (Schwab et al, 2006;Molnar et al, 2009). The use of two BsaI sites in this configuration has been adapted from the Golden Gate cloning method (Engler et al, 2008) and was used in other amiRNA cloning methods (Chen et al, 2009;Zhou et al, 2013). BsaI digestion of the B/c vector and subsequent ligation of the amiRNA oligonucleotide insert can be done in separate reactions or combined in a single 5-min reaction (Supplemental Protocol S1).…”
Section: Atmir390a-based Vectorsmentioning
confidence: 99%
“…Recently, in animals and plants, endogenous miRNA precursors have been used to develop artificial miRNA technology. This has led to the generation of targetspecific miRNA for gene silencing (Schwab et al, 2006;Molnar et al, 2009;Zhou et al, 2013).…”
Section: Introductionmentioning
confidence: 99%