gRNA transcription by pol ii promoter

is it possible to use pol ii promoters for better control over gRNA (or precrRNA) transcription? Did someone try this before and if so what are the tricks and caveats (e.g specific UTRs, removing the PolyA with a hammerhead, using only intronic sequences etc.)?


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Navy Gorgonalmost 9 years ago

They have tried that in Ren et al. PNAS 2013 for CRISPR in Drosophila and it did not work at all.

Snow Kappaover 8 years ago

we have just published a paper in which we demonstrate the expression of multiple gRNA species from a single Pol II promoter in human cells:

Multiplexed and Programmable Regulation of Gene Networks with an Integrated RNA and CRISPR/Cas Toolkit in Human Cells

http://www.sciencedirect.com/science/article/pii/S1097276514003554

Seashell Griffinover 3 years ago

Can the HH ribozyme allow extra nucleotides on the 5' end which are not used for secondary structure?

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