Technique / Molecular Biology / RNAi siRNA gene silencing

shRNA short hairpin RNA

shRNA short hairpin RNA

shRNA

shRNA (small hairpin RNA) constructs is an important gene silencing tool for long term and stable gene expression knockdown. Design of shRNA constructs is often very useful for silencing expression of slow turnover genes. There are several commercially available plasmid and lentiviral vectors for shRNA construction. Some vendors for shRNA constructions are listed here.

shRNA from sigma

TRC Protocols, vector maps (MIT)
Glycerol stock plasmid prep
Lentivirus production and infection
pLKO1 map and sequence

TRC lentiviral shRNA library (open biosystems)
lentiviral shRNA libraries targeting 15,000 human (TRC-Hs 1.0) and 15,000 mouse (TRC-Mm1.0)

shRNA construction kits (Clontech)
Pool of functionally validated shRNA vectors for gene-specific knockdown. Both plasmid and virial delivery are included in the kit.

SMARTvector shRNA Lentivirus(ThermoFisher scientific, Dhamracon)
Smartvector shRNA kit takes advandage of miRNA scaffold-specific attributes of lentivirus to knock down gene expression via endogenous RNAi pathway.

Protocols for shRNA construction

shRNA lentivirus design and cloning (open access protocol from cold spring harbor lab).
This protocol was adapted from "Development of Lentiviral Vectors Expressing siRNA," Chapter 3, in Gene Transfer: Delivery and Expression of DNA and RNA (eds. Friedmann and Rossi). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2007.

This protocol combines the specificity of small interfering RNA (siRNA)-mediated silencing cassettes with the versatility of lentiviral vectors to stably transduce a wide range of cell types.

shRNA oligo designer (online shRNA design tool)