Part:BBa_K1733000:Experience

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Applications of BBa_K1733000

We tested this orf7 RDF in our mammalian systems with a reporter that contained an unexpressed fluorescent protein flanked by AttL and AttR recombination sites. The RDF recognized these AttL and AttR recombination sites and, when both the RDF and its corresponding integrase, TP901-1, the proteins will flip the sequence in between so that the fluorescent protein is then expressed.

This part is used in conjunction with TP901-1. Using these two proteins together can create a bistable switch. TP901-1 recognizes AttB and AttP recombination sites, and flips the sequence that these recombination sites flank. The recombination sites then become AttL and AttR, which means the sequence cannot be flipped by TP901-1 itself. The addition of the orf7 RDF will flip this sequence again, because the RDF recognizes the AttL and AttR sites. After this flipping reaction, the recombination sites once again become AttB and AttP.

Using these two proteins, it is possible to knock out or knock in a certain gene of interest by inverting the sequence and rendering it gibberish.

User Reviews

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