Z**********g
发帖数: 222 | 1 本人对mir是外行,一些疑问想请教大家.
1.mir的binding site刚开始认为在靶基因mRNA的3'UTR,但后来又有报道表明mir也可能
binding在5'UTR或coding region.那这样的话,目前常用的luciferase reporter方法是
否适用于binding site在5'UTR或coding region的情况?我看文献大多是只检测3'UTR的
情况.
2.近期David Bartel的一篇Nature(2010 Aug 12;466(7308):835-40)说哺乳动物的mir
主要通过降低靶基因的mRNA level (destablization)而起作用的,而不是之前认为的干
扰mRNA翻译.那这样的话,luciferase reporter方法还适用否?或者说,是否应该和siRNA
一样,应该灵敏的检测到靶基因mRNA level的变化?
请牛人们帮忙解惑哦. | l******e
发帖数: 427 | 2 1. pairing is widespread to occur at the 3'utr. if you have candidates,
focus on 3'utr first. donot get distracted by sporadic publications (i don'
t mean that they are false; i mean they are not representative.)
2. it is not a new story. once again, focus on protein levels. no matter
how miR might operate, the effect on protein levels will reflect the effect.
.. (I used to like david bartel very much, but now i feel more and more
disappointed by his work, anyway...) | Z**********g
发帖数: 222 | 3 Good points.I've got it.
don'
effect.
【在 l******e 的大作中提到】
: 1. pairing is widespread to occur at the 3'utr. if you have candidates,
: focus on 3'utr first. donot get distracted by sporadic publications (i don'
: t mean that they are false; i mean they are not representative.)
: 2. it is not a new story. once again, focus on protein levels. no matter
: how miR might operate, the effect on protein levels will reflect the effect.
: .. (I used to like david bartel very much, but now i feel more and more
: disappointed by his work, anyway...)
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