Advantages of Using pRNAi
 Figure 1. Structure map of pRNAi vector system. |
| Biosettia’s pRNAi Vector is Ready-To-Use |
| -No restriction digestion or vector purification is required |
| Reduced Cloning Complexity |
| -Our single-strand DNA oligo encoding shRNA sequence is a perfect palindrome, thus the same (two) palindromic oligos can anneal to each other to form the final double-strand oligo. This eliminates the need to mix and anneal two different DNA oligos and reduces operational mistakes during the cloning process. The overall chance of mutations introduced during DNA oligo synthesis is therefore reduced by 50% (Fig. 2A). |
| Low Cost |
| –Only one DNA oligo is required to generate an shRNA expression clone. This setup is the most economical and is distinguishable from all the other shRNA vectors commonly used in academia and industry, for which two different oligos are needed to make a shRNA expression clone. Also, the length of the DNA oligo used for pRNAi (52-56 nt) is shorter than the length of DNA oligos required for most of the other commercially available shRNA vectors. |
| Highly efficient, Low Background shRNA Cloning |
| –5’-AAAA overhangs on annealed double-strand oligo can only be ligated to the 5’-TTTT overhangs on both ends of the linear pRNAi vector. Unlike the overhangs generated by restriction digestion in other available shRNA expression vectors, these 5’-TTTT overhangs in pRNAi will not self-ligate. As a result, the ligation of the double-strand oligo into the pRNAi vector is highly efficient, and the contamination of empty, self-ligated pRNAi vectors is greatly reduced (Fig. 2B). |
| Three Promoters to Choose From |
| -Biosettia’s shRNAs can be expressed from human H1 (H1), human U6 (hU6), or mouse U6 (mU6) promoters in the pRNAi system. You are able to choose the best suitable the pol III promoter for their model system. |
| Long Term Suppression |
| -Biosettia’s pRNAi vectors encode antibiotics resistance markers for puromycin (Puro), hygromycin (Hyg), neomycin (Neo), or blasticidin (Bsd), allowing you to generate stable cell lines with long-term suppression of your genes. |