3 replies to this topic

[Jon Moulton]

Nicoli S, Standley C, Walker P, Hurlstone A, Fogarty KE, Lawson ND. MicroRNA-mediated integration of haemodynamics and Vegf signalling during angiogenesis.  Nature. 2010 Apr 4. [Epub ahead of print]

Edited by Jon Moulton, 13 April 2010 - 10:59 AM.

[luc]

Jon Moulton, on Apr 13 2010, 10:53 PM, said:

Nicoli S, Standley C, Walker P, Hurlstone A, Fogarty KE, Lawson ND. MicroRNA-mediated integration of haemodynamics and Vegf signalling during angiogenesis.  Nature. 2010 Apr 4. [Epub ahead of print]

hi
I have read this paper too . But I have a question about this paper. can you help me ?
  
To construct the microrna expression vector they constructed a middle
entry (pME) clone containing a 753 bp fragment of the zebrafish EF1alpha gene .

why can not just placed the pri-mirna directly behind the promoter?

thank you

[Jon Moulton]

luc, on May 21 2010, 06:07 AM, said:

To construct the microrna expression vector they constructed a middle
entry (pME) clone containing a 753 bp fragment of the zebrafish EF1alpha gene .

why can not just placed the pri-mirna directly behind the promoter?

thank you

This is a guess, Luc - I've not made a similar construct.  However, it makes sense to me that the pri-miRNA must be presented with some length of RNA extending out past the 3' and 5' Drosha cleavage sites in order to be recognized by Drosha for the first cleavage of miRNA maturation.  If the construct were inserted directly downstream of the promoter then it could be that the length of RNA to the 5' of the Drosha site would be too short and Drosha would not bind well.  I suppose if the insert included enough 5' sequence this wouldn't be a concern, but perhaps they were working with a short insert.

[ljchen]

Jon Moulton, on Apr 13 2010, 10:53 PM, said:

Nicoli S, Standley C, Walker P, Hurlstone A, Fogarty KE, Lawson ND. MicroRNA-mediated integration of haemodynamics and Vegf signalling during angiogenesis.  Nature. 2010 Apr 4. [Epub ahead of print]

This is a nice paper!! thank you!!