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How to clone two mir-30 based shRNAs into the same vector and express them from

mir-30 shRNA multiple shRNA cloning

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#1 5280

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Posted 01 August 2015 - 02:52 PM

Hi, 

 

I want express two mir-30 based shRNAs from a single transcript. I plan to modify the TRIPZ inducible system as following:

 

PolII inducible promoter - RFP - shRNA1 - shRNA2 (Clone in second shRNA). 

 

Do I need a specific distance or sequence between shRNA 1 and 2? I assume RFP-shRNA1-shRNA2 will expressed as a single transcript. RFP will then be translated and shRNA1 and 2 will be processed from the transcript. Correct? 

 

I cannot find any literature about this so please let me know if there is a good paper or if you have suggestions for the cloning. 

 

Thanks!







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