n*******l
发帖数: 75 | 1 请教构建lentivirus质粒的大拿，菜鸟求教
我想沉默pc12细胞的一个基因。查到某公司有公布的基因的沉默序列，在pGFP-shlenti
上。我想构建到lvthm上
问题是我们实验室构建的模版都是（21nt+9bp的loop+21nt的反向互补）在lVTHM上。公
司公布的是（29nt+7bp+29nt的反向互补）
。直接把用（29nt+9bp的loop+29nt的反向互
补）可以直接构建到lvthm上 影响沉默效率么，另外，loop的长短会影响沉默效率么。
万分感谢。 |
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r***e
发帖数: 2539 | 2 长度没问题（19-29都有），序列才是关键。最近在做一个22nt的序列，我一不小心漏了
5'一个nt，成了21nt竟然一点都不行。但并不是说21nt长度不行。（有点拗口，但就是
这个意思）
shlenti |
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c*********t
发帖数: 340 | 3 ffff6699 is right
I just had an exam on this question:)
siRNA is exogeneous,about 21nt, usually completely complementary
miRNA is endogeneous, about 70nt, processed to give smaller, mature miRNA of
approximately the same size of that of siRNA; usually not 100%
complementary. Could result in mRNA degradation, inhibition of translation,
etc.. For translation inhibition it binds to 3'UTR most of the times but
some miRNA binds to 5'UTR or intronic regions too
This was what i wrote down and got full |
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f****6
发帖数: 1 | 4 本人发现target sequence 19nt--21nt的shRNA 的效果不好。 |
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F*K
发帖数: 608 | 5 Hi all,
I plan to rescue knockdown of a mouse gene with human cDNA.
However, the human gene only mismatches the 21nt shRNA on #2, 17 and 20
positions. I am wondering if it is ok for rescue?
thanks |
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