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Custom-made shRNA Lentivirus Plasmids (SH2001)

Availability: In stock


Quick Overview

For each target, ATCGbio Life Technology Inc.provides two shRNA lentivirus plasmid with a negative control. The plasmids we provide are ready to be amplified. The latest our data showed 90% of shRNA vector represses target mRNA more than 70%.

Customer can choose the region of knocking-down (CDS or 3'-UTR), reporter co-expression (GFP, RFP or iRFP) and antobiotics resistance (puromycin, hygromycin, blasticidin or zeocin).

The plasmids package contains:
1. Negative-control shRNA lentivirus plasmid 20µl (~50ng/µl)
2. Custom-made shRNA lentivirus plasmid   20µl (~50ng/µl) x 2

In 4-5 weeks, lentivirus plasmids will be shipped in 0.5 ml tube, and users are free to amplify. Upon receiving the tubes, keep it at 4 °C till amplification.


Order Instruction

Please submit the information into the following box for order:
1. Please do not submit conventional name of protein (e.g. SIRT1).
2. Submit an official symbol and gene ID or Accession number (If specific isoform targeting shRNA is required, please submit isoforms' number). Please do not submit conventional name of protein (e.g. SIRT1). Go NCBI gene website (http://www.ncbi.nim.nih.gov/gene) and look for gene ID and official symbol.
  For example, in case of human sirtuin 1:
                    The gene ID is 23411 and official symbol is SIRT1.
3. Region of knocking-down (CDS or 3'-UTR)
4. Reporter co-expression: GFP, RFP or iRFP
5. Antibiotics resistance: puromycin, hygromycin, blasticidin or zeocin
If you don't sepcify for 3, 4, and 5, we'll offer you a viral particle expressing shRNA for target gene at CDS or 3'-UTR, GFP co-expression and hygromycin resistance.
If your target genes are 2 or more, please enter the number of your target genes (not the number of the isoforms) in the "Qty" box before you click "ADD TO CART".
6. We may contact after feasibility evaluation.

Order Form

Custom-made shRNA Lentivirus Plasmids (SH2001)

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ATCGbio Life Technology Inc. provides function-validated shRNA (short-hairpin RNA) in lentivirus plasmid.User can amplify plasmid and create lentivirus. We recommend to use our VP-Easy Kit (VP1001) to amplify the plasmid and pLenti™ Lentivirus Production Kit (LT1001) to produce lentivirus. Once virus is created, infected cells express shRNA (driven by human U6 promoter) to knock-down specific mRNA* or non-coding RNA.  Our shRNAs are designed by our proprietary design scheme having the following features.  

1. High specificity (at least 4 mismatches in 19 nt stem)

2. High target accessibility (calculated based on RNA structure)

3. High loading efficiency of guide strand (high 5’~3’ energy disparity)

4. Accurate position cut and loading to RISC (newly developed loop structure)

5. Based on above features, our shRNA plasmids have highly effective

6. The basal construct is based on the third generation design.

7. It expresses GFP and hygromycine resistance gene product.

* shRNA works on mRNA which is not always correlated with protein levels. In case of protein expression is largely regulated by protein degradation (e.g. ubiquitination-proteasome system), mRNA levels may not be critical point to regulate protein expression. However, in case of this issue, it is recommended to re-infect cell 2-3 more times if the cell condition allows longer period of culture especially cancer cells.

The following figure is the results of experiments proves our shRNA performance

(One shRNA sequence was selected for one target in each design)

         shRNA-lentivirus-performance                   Non-target   Gene-specific
               shRNA           shRNA
• shRNA Lentivirus were created to target 10 transcription factors expressed in HepG2 cells.
• 64 hrs after infection without antibiotic selection, the target gene mRNAs were down to 28.8% in average of 10 shRNA target (p<0.0001 by paired t-test). 9 out of 10 design shRNA were able to suppress more than 70%, and only one out of 10 shRNA suppressed more modestly about 60%.
• Suppression will be greater, when the cells would be cultured longer time with antibiotics selection.

Ten transcription factors expressed in HepG2 cells were selected for this experiment. The lentivirus plasmids expressing shRNA were made for each target gene using our propriety design scheme (One shRNA sequence was selected for one target in each design). Lentivirus was created in 12-well plates using our pLenti™ Lentivirus Production Kit (LT1001), which produces a high titer stable lentivirus solution in 48hrs. HepG2 cells were grown in 96-well palate and infected with each virus including negative control shRNA by adding 50µl crude virus solution into HepG2 cells cultured with100µl culture medium for overnight(16hrs.). The cells were incubated further 48hrs., and harvested for Direct RT real-time PCR.  As shown in the right figure, 64 hrs. after infection, target gene mRNAs were down to 28.8% in average of 10 shRNA target. 9 out of 10 design shRNA were able to suppress more than 70%, and only one out of 10 shRNA suppressed more modestly about 60%.

The experiment results of lentivirus production and infection (lentivirus produced by pLenti™ Lentivirus Production Kit (LT1001) and our shRNA Lentivirus Plasmids (SH2001)):

Fig.A and Fig.B (magnification 100x): GFP expressing lentivirus was created with two different medium

    Conventional DMEM+10% FBS                              Our Lentivirus Production Medium + 2% FBS

 Lentivirus-infect293T-cell(ordernary-medium-cell-culture) Fig.A  

       Lentivirus-infect293T-cell(our-Production-Medium-culture) Fig.B       

50µl of 3ml crude lentivirus with medium was directly used to infect 293T cells (without polybrene) grown in a separate 6 well plate. 48 hours later the expression of GFP was assessed.

Fig. C and Fig. D (magnification 100x): shRNA experssion lentivurs was produced by the pLenti™ Lentivirus Production Kit (LT1001); and infected to HUVEC (Human Umbilical Vein Endothelial Cells).

Primary human umbilical vein endothelial cells (HUVEC) growing in 100mm dishes were infected by lentivirus shRNA negative control co-expressing with GFP. GFP expression were observed 3 days after infection.

Lentivirus-infection-in-human-endothelial-cells1-3br.jpg Fig.C

    Lentivirus-infection-in-human-endothelial-cells1-3fl.jpg Fig.D
   Fig.E (magnification 100x)            Lentivirus-in-vivo-example(fat-injection)                          

Fig.E: In vivo transduction sample

50 µl of purified lentivirus with our kit was injected into a mouse subcutaneous adipose tissue (without polybrene). After three days, the tissue was harvested and observed under fluorescence microscope and bright field.



 1. Y. Ido, et. al. PLoS ONE, 2012 Apr 07( 4): e35092
 2. Lan F, et. al. J Biol Chem. 2008 Oct 10;283(41):27628-35.

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