In my lab we have miR-145 cloned into pCMV-MIR vector which is specific for mouse according to vendors website they have different plasmids for for human Mouse Rat. Mature sequence of the miR-145 is same in Human and mouse but precursor miRNA sequence is different. So can I use mouse specific miR-145 vector to study the effect of this miRNA in human cell lines.
I presume your vector encodes precursor sequence for miRNA over expression. In theory, minor sequence differences in the non processed part of pre-miRNAs should not make a difference to the final product being generated. You can give it a try. I have personally used clones generated way back in 2006 for mouse in human cells. The group even published the work. When I asked about it, their reply was that they didn't know much about the difference it would make back then. But publishing it today might raise eyebrows.If its a miRNA sponging/inhibitor construct you can use it.
vector have insert around 700 bp I think it encodes primary miRNA sequence. Kindly see the attachment
Looks like the human version is not identical. Mouse and rat are 100% identical.So I would not use it in this case.
http://blast.ncbi.nlm.nih.gov/Blast.cgi?CMD=Web&PAGE_TYPE=BlastSearch&VIEW_SEARCH=on&UNIQ_SEARCH_NAME=A_SearchOptions_1ahIPy_4DF_du1yfdCaGaI_23turP_H2Xwu
Thank you so much Ratnakar for your answer.I think I have to buy new vector specific for Human.
Hello Nishant,
Our concept of miRNA and its role in diseases took a major turn when we found that alien miRNAs can also survive in our system. If you have the time kindly read the attached manuscript. It will interest you.
Warm regards,
Venk
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