What is shRNAmir?

shRNAmir are expressed as primary-miRNA (pri-miRNA) transcripts. These constructs were created by redesigning the well- studied human miRNA, miR-30, to express an artificial siRNA/miRNA. The stem of the primary miR-30 transcript was replaced with gene-specific duplexes for different target genes (Figure 2).

How long does it take shRNA to work?

Gene silencing resulting from siRNA can be assessed as early as 24 hours post-transfection. The effect most often will last from 5–7 days. However, the duration and level of knockdown are dependent on the cell type and concentration of siRNA.

What is shRNA construct?

Small hairpin RNA (shRNA) and lentiviral vector RNA interference has been used as a research tool to control the expression of specific genes in numerous experimental organisms and has potential as a therapeutic strategy to reduce the expression of problem genes[1,2].

How does shRNA knockdown work?

shRNA molecules are processed within the cell to form siRNA which in turn knock down gene expression. The benefit of shRNA is that they can be incorporated into plasmid vectors and integrated into genomic DNA for longer-term or stable expression, and thus longer knockdown of the target mRNA.

How are shRNA designed?

Kay Lab siRNA/shRNA/Oligo Optimal Design shRNA (short hairpin RNAs) are artificial constructs that can be expressed endogenously. These expressed hairpins fold to form dsRNA, and Drosha and Dicer then act on these hairpins to create mature sequence, used by the RISC complex to target genes.

How is shRNA made?

shRNAs are synthesized in the nucleus of transfected/transduced cells and form hairpin structures that consist of a stem region of paired antisense and sense strands connected by unpaired nucleotides that make up a loop (Figure 1b and 1c). They are converted into siRNAs by the same RNAi machinery that processes miRNAs.