We provide suggestions for designing shRNA targets and controls, a protocol for sequencing through the secondary structure of the shRNA hairpin structure, and protocols for packaging and delivery of shRNA lentiviral particles. Using real-time PCR and functional assays we demonstrate the successful knockdown of ASC, an inflammatory adaptor molecule.

Short hairpin RNA (shRNA) sequences are usually encoded in a DNA vector that can be introduced into cells via plasmid transfection or viral transduction. shRNA molecules can be divided into two main categories based on their designs: …

A short hairpin RNA or small hairpin RNA (shRNA /Hairpin Vector) is an artificial RNA molecule with a tight hairpin turn that can be used to silence target gene expression via RNA interference (RNAi). [1][2] Expression of shRNA in cells is typically accomplished by delivery of plasmids or through viral or bacterial vectors. shRNA is an ...

What is sh RNA? Vector-based short hairpin RNA (shRNA) is a type of RNA interference (RNAi) technology leveraged to study the function of unknown genes. RNAi works by by silencing gene function to allow for the examination of the affected processes.

2006年1月5日 · Short hairpin RNA (shRNA) encoded within an expression vector has proven an effective means of harnessing the RNA interference (RNAi) pathway in mammalian cells. A survey of the literature revealed that shRNA vector construction can be hindered by high mutation rates and the ensuing sequencing is often problematic.

Here, based on current understanding of the miRNA maturation pathway, we discuss the principles of different shRNA design (pre-miRNA-like, pri-miRNA-like and Ago-shRNA) with an emphasis on the RNA structure.

The Ago-shRNA pathway is Dicer-independent and critically involved in the biogenesis of miR-451. The m 7 G-capped pre-miRNA pathway bypasses Drosha processing and can employ XPO1 to subsequently export the pre-miRNA from the nucleus to the cytoplasm.

2012年1月1日 · Since the first application of RNA interference (RNAi) in mammalian cells, the expression of short hairpin RNAs (shRNAs) for targeted gene silencing has become a benchmark technology. Using plasmid and viral vectoring systems, the transcription of shRNA precursors that are effectively processed by the RNAi pathway can lead to potent gene knockdown.

Short hairpin RNA (shRNA) is an alternative way to prepare siRNA sequences for delivery to cells that can be expressed in situ from plasmid DNA (pDNA) or from virus-derived constructs. The effectiveness of shRNA was first reported by Paddison and Hannon in 2002 [48].

Vector-based short hairpin RNA (shRNA) is a type of RNA interference (RNAi) technology leveraged to study the function of unknown genes. RNAi works by by silencing gene function to allow for the examination of the affected processes.