ShRNA expression cloning

OmicsLink ™ Characteristics of shRNA expression clones
All shRNA target sequences were selected through accurate algorithmic calculations. According to different genes, the stem length of shRNA ranges from 19 to 29 base pairs. These guarantee high expression inhibition efficiency and minimal off target effect after shRNA cloning expression.
The vectors used for cloning shRNA include lentiviral vectors and mammalian cell based vectors, each with different promoters and tracking marker genes to choose from.
The green fluorescent tracking marker gene (eGFP) carried in the vector can be used to monitor the transfection efficiency of plasmid vectors and the infection efficiency of transduced viruses.
The carrier structure contains screening markers for stable cell lines (puromycin) and green fluorescent tracking marker genes (EGFP), which enable the carrier to be used for both stable inhibition and transient inhibition studies.
Using a lentiviral vector system, shRNA can be transduced into conventional dividing cells, as well as target cell lines that do not divide and are difficult to transfect.
The sequences of various parts of the vector have been sequenced, including the promoter, sense and antisense strands encoding shRNA, hairpin structure, terminator, and other linker sequences.

OmicsLink ™ The application
The inhibitory effect of downregulated shRNA cloning on gene expression of a single gene can be compared and studied with a negative control clone containing an irregular sequence.
Our company's human gene ORF clones have been classified according to different signaling pathways, metabolism, disease pathways and correlations, gene families, etc. By arranging shRNA clones with known identical pathways in 96 well or 384 well plates, the function of a group of genes in a certain signaling pathway can be studied at once. At present, our company can provide shRNA clones targeting all kinase genes in humans, and has classified them according to the functions of different kinases.
ORF expression cloning confirms that our company already has ORF clones of shRNA corresponding to human target genes, which can be installed in various expression vectors with different tags and tracking marker genes. These normal ORF expression clones and the same ORF expression clones with silencing mutations in the shRNA target sequence can be used for shRNA confirmation studies and gene function rescue studies.

Product Assurance

The expression frame has been fully sequenced, including promoter, sense and antisense target sequences, hairpin structure, termination signal, and other linker sequences. For the human kinase group shRNA clones constructed into the psi-sH1 vector, we provide 1-3 clones for each gene, and all clones have been confirmed by AP colorimetry to have an expression inhibition level of over 70% on the target gene. For constructing shRNA clones into lentiviral and mammalian vectors, we provide 4 clones per gene. We guarantee that at least one clone achieves an inhibition level of 70% or more on the expression of the target gene at the mRNA level, provided that the detection method mentioned earlier is used and the experiment is conducted strictly in accordance with the operating procedures and precautions provided in the instructions. If our company verifies that none of the four have the expected expression inhibition level, we promise to replace 1-4 shRNA clones free of charge.


*This product is for research purposes only and is not intended for clinical diagnosis