How many bp should be left at 5' and 3' flanking regions needed for miRN - miRNA (Mar/23/2009 )

I am anxious to know.


How many bp should be left at 5' and 3' flanking regions to constucting a miRNA vector?


Some people left 600bp, some people left 100 or 200 bp. How many should be enogh for expressing miRNA?


Is there any article about this?

Thank?

100-200 bp flanking the precursor miRNA should be fine.

Thank you pcrman.

If a longer bp was remained, any problem will happen?
Thanks.

pcrman on Mar 24 2009, 07:01 AM said:

I tried 30 and 60.....30 was more than enough!!
but maybe i was just lucky.
Fiz

I have cloned and generated over 600 human lentiviral miRNA collection. There are 100bp flanking sequences on both sides of pre-miRNA and they work great.