shRNA-expressing constructs are genetic constructs that encode short hairpin RNAs (shRNAs) for the purpose of inducing RNA interference (RNAi) in cells. These constructs are widely used in research to study gene function and to develop new therapies for a variety of diseases.
It typically consist of a promoter, a shRNA sequence, and a termination sequence. The promoter drives the expression of the shRNA, which is typically designed to target a specific messenger RNA (mRNA) molecule. The termination sequence ensures that the shRNA is processed correctly and that the correct size of the shRNA is produced.
There are several types of shRNA-expressing constructs, including plasmids, viral vectors (such as lentivirus or retrovirus), and bacterial artificial chromosomes (BACs). These constructs can be delivered to cells using a variety of methods, including transfection, electroporation, and viral transduction.
They have several advantages over other RNAi methods, such as siRNA delivery or CRISPR-Cas9 gene editing. These advantages include:
- Stable gene knockdown: provide stable and long-term knockdown of gene expression, allowing for extended studies of gene function.
- Low off-target effects: can be designed to target specific genes, reducing the risk of off-target effects.
- Efficient screening: can be used for high-throughput screening of potential drug targets and therapeutic agents.
Those constructs are used in a wide range of applications, including:
- Gene function studies: they are used to study the function of specific genes in cells and organisms.
- Therapeutic development: they can be used to develop RNA-based therapies for the treatment of diseases such as cancer and viral infections.
- Disease modeling: they can be used to create models of disease by inhibiting the expression of genes associated with specific diseases.
Overall, shRNA-expressing constructs are a powerful tool for studying gene function and developing new therapies, and they have the potential to accelerate the discovery of new treatments for a wide range of diseases.