shRNA Kits

Pre-made shRNA Lentivirus Plasmids (SH1001)

AbDia Technologies Inc. provides very effective or function-validated shRNA (short-hairpin RNA) in lentivirus plasmid.  User can amplify plasmid and create lentivirus. We recommend using our VP-Easy^TM 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. 

* 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.

Pre-made shRNA Expression Lentivirus Particles (SH3001)

The shRNA Lentivirus particles from AbDia Technologies 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 AbDia Technologies are created base on safer 3rd generation recombinant lentivirus vector through our new shRNA design system; and produced by our own pLenti^TMLentivirus 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.

Table.1.

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

References

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