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shRNA Products

Pre-made shRNA Lentivirus Plasmids

Custom-made shRNA Lentivirus Plasmids

Pre-made shRNA expression lentivirus Particle

Custom-made shRNA expression lentivirus Particle

Lentivirus Plasmids

ATCGbio Life Technology Inc. provides very effective or function-validated shRNA (short-hairpin RNA) in lentivirus plasmid.  User can amplify plasmid and create lentivirus. We recommend to use our VP-EasyTM Kit to amplify the plasmid and Lentivirus Production Kit 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. 

We provide only up to two plasmids for each target. The effect of knocking down is guaranteed (>70%). If user needs more shRNA sequences for same target, please CONTACT US, or email at info@atcgbio.com.

Our shRNAs are designed by our proprietary design scheme having the following features.  

1. High specificity (there are at least 4 mismatches to non-target mNRA)

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. In such a case, measuring mRNA is recommended to confirm the function.

Lentivirus Particles

The shRNA Lentivirus particles from ATCGbio Life Technology are recombinant shRNA expression lentivirus particle solution to be ready to infect directly for knocking-down gene of interset. It’s specifically designed for scientists who don’t have time or don’t prefer to make lentivirus by themselves. It’s the easiest way to knockdown gene of interest.

shRNA Lentivirus particles from ATCGbio Life Technology are created base on safer 3rd. generation recombinant lentivirus vector through our new shRNA design system; and produced by our own pLentiTM Lentivirus Production Kit (LT1001). Virus expresses shRNA for target gene, GFP protein and hygromycine resistance gene product. It is suitable for both cultured cells and in vivo injection to small animals.  Lentivirus solution is DMEM based medium containing 2% FBS and our lentivirus precipitation/purification solution. It does not contain any chemicals used to boost lentivirus titers such as chloroquin, acetate or nicotines. The solution can be used directly to the cells growing in DMEM base medium without further precipitation/purification. There is no visible viral toxicities (e.g. change in cell morphology or/and cell detachment from culture plate) if the viral solution added up to a half amount of cell culture medium as described in table 1. below. Some special cells such as keratinocytes may be used to grow with special medium which is incompatible with DMEM or FBS. In such a case, it is recommended to precipitate/purify lentivirus and reconstitute it with storage solution as described in User Manual.

shRNA-Lenti particle solutions from ATCGbio Life technology include two products of categories: Pre-made shRNA-Lenti particle solution (SH3001) and Custom-made shRNA-Lenti particle solution (SH4001).The highlight features of the products are:
  1. Most advanced shRNA design.Near 100% success rate (Fig.1).
  2. High infectious efficiency (Fig.2), up to 0.9 x 108 TU/ml.
  3. Co-express GFP and hygromycine resistance gene.
  4. Just add and incubate overnight (16 hrs).
  5. No cytotoxicity. Small animal injectable without immonoreactive symptoms.
  6. Unlike siRNA, transduced cells show knock-down effects even in divided cells.
  7. No need to add polybrene. Near 100% transduction is possible.
  8. Cell-line can be made in a week.
  9. Suitable for both primary cells and transformed cells.


                           Size of Culture          Culture Medium         Lentiviral Solution      
  1 well of 6-well plate 1.0ml 0.5ml
  1 well of 12-well plate 0.5ml 0.25ml
  1 well of 24-well plate 0.25ml 0.125ml


Fig.1. HepG2 cells grown in 100µl MEM+10% FBS on the 96-well plate were infected with 50µl shRNA lentivirus solution 48hrs.
Near 100% transduction.
Fig.2. 64hrs after infection without antibiotic selection, the target gene mRNAs were down to 28.8% in average of 10 shRNA targets in HepG2 cells.
More details are described below.

shRNA-lentivirus-performance                                 Non-target   Gene-specific                           shRNA          shRNA

As shown in Figure 2. 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 pLentiTM Lentivirus Production kit, 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 targets (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.
The experiment results of lentivirus production and infection (lentivirus produced by pLenti™ Lentivirus Production Kit (LT1001) and our shRNA Lentivirus Plasmids SH2001 or SH1001 ):

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
  1. Lan F, et al.  J Biol Chem. 2008 Oct 10;283(41):27628-35


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