This research article investigates the use of artificial microRNAs (amiRNAs) to achieve highly specific gene silencing in Arabidopsis thaliana. The authors designed amiRNAs to target individual genes or groups of endogenous genes, similar to natural plant microRNAs (miRNAs). They found that amiRNAs were as effective as natural miRNAs in silencing target genes, with high specificity and limited off-target effects. The study demonstrated that known parameters of target selection for natural miRNAs accurately predicted the targets of amiRNAs, indicating that the plant silencing machinery acts with higher specificity than its animal counterpart. Additionally, amiRNAs were shown to be effective when expressed under tissue-specific or inducible promoters, with limited nonautonomous effects. The design principles for amiRNAs have been generalized and integrated into a web-based tool for automated design. The research highlights the potential of amiRNAs as a powerful tool for specific gene silencing in plants, particularly for targeting multiple related genes.This research article investigates the use of artificial microRNAs (amiRNAs) to achieve highly specific gene silencing in Arabidopsis thaliana. The authors designed amiRNAs to target individual genes or groups of endogenous genes, similar to natural plant microRNAs (miRNAs). They found that amiRNAs were as effective as natural miRNAs in silencing target genes, with high specificity and limited off-target effects. The study demonstrated that known parameters of target selection for natural miRNAs accurately predicted the targets of amiRNAs, indicating that the plant silencing machinery acts with higher specificity than its animal counterpart. Additionally, amiRNAs were shown to be effective when expressed under tissue-specific or inducible promoters, with limited nonautonomous effects. The design principles for amiRNAs have been generalized and integrated into a web-based tool for automated design. The research highlights the potential of amiRNAs as a powerful tool for specific gene silencing in plants, particularly for targeting multiple related genes.