HuSH-29 shRNA Vectors

All of the plasmids within the HuSH shRNA collection are cloned into OriGene's non-proprietary pRS, pGFP-V-RS or pRFP-C-RS vector, allowing both transient and stable transfection, as well as stable delivery of the shRNA expression cassette into host cells via a replication-deficient retrovirus. The pGFP-V-RS and pRFP-C-RS vector offers additional feature of turbo-GFP and turbo-RFP expression that facilitates easy monitoring of single or dual-gene transfection. (User Manual)

Features pRS pGFP-V-RS pRFP-C-RS pGFP-B-RS Fluorescent Label N/A Green Red Green Selection Marker in E.coli Ampicillin Kanamycin Chloramphenicol Kanamycin Selection Marker for Mammalian Puromycin Puromycin Puromycin Blasticidin Transient or Stable Transfection Yes Retroviral Packaging Yes Download Sequence Download Datasheet Price / Amount / Delivery $150 / 5ug / Immediate

pGFP-V-RS vector: The HuSH pGFP-V-RS plasmid vector (Figure 1) contains both 5’ and 3’ LTRs of Moloney murine leukemia virus (MMLV) that flank the puromycin marker and the U6-shRNA expression cassette. Upon transient transfection of the plasmids into a packaging cell line, replication deficient viruses can be obtained and used to infect target cells. The puromycin-N-acetyl transferase gene and Kanamycin gene provide selection of antibiotics puromycin and kanamycin, respectively. There is an integrated turboGFP element driven by a cMV promoter to readily verify transfection efficiency. For our RNAi products, the shRNA expression cassette consists of a 29 bp target gene specific sequence, a 7 bp loop, and another 29 bp reverse complementary sequence, all under human U6 promoter. A termination sequence (TTTTTT) is located immediately downstream of the second 29 bp reverse complementary sequence to terminate the transcription by RNA Pol III. The gene-specific shRNA cassette is sequence-verified to ensure its match to the target gene.
pRFP-C-RS vector: The HuSH pRFP-C-RS plasmid vector (Figure 2) was created with an integrated turboRFP element to readily verify transfection efficiency. It incorporates both a chloramphenicol and puromycin resistance elements for greater selection capabilities. The pRFP-C-RS plasmid is also ideal for monitoring the dual-gene knockdown experiments when used alongside pGFP-V-RS expression plasmid.
pRS vector: The HuSH pRS plasmid vector (Figure 3) incorporates all the elements as above two vectors with the absence of a fluorescence marker. The bacterial selection marker for pRS vector is ampicillin and the mammalian selection can still be achieved with puromycin.
pGFP-B-RS vector: The HuSH pGFP-B-RS plasmid vector (Figure 4) offers the same elements as pGFP-V-RS vector with an exception of an alternative mammalian selection marker. The substitute feature of using blasticidin selection instead of puromycin, which is available in all of our retroviral vectors assists researcher in creating stable cell lines in double knockdown experiment. Visit stable cell lines in double knockdown experiments to learn more..

Additional control vectors are available and can be purchased seperately.