Hi, I would like to functionally validate a -not seen before- 3 bp deletion I found on a 5' UTR of a knoen disease causing gene. I looked at the UCSC browsers tracks for conservation and regulation and it seems like this region is both well conserved and has a lot of regulation signals associated with it. Looking at gene expression by RT-PCR looks like the best option but what other next steps would be also valuable on the path to functionally validate this mutation?
Thanks
1 answer
To complete the validation I will suggest an in-vivo experiment using transfected cell-lines (the expression can be quantified using RT-PCR or fusion your 5'UTR to a reporter like GFP), testing: 1) the 5' UTR with the 3bp deletion, 2) the wild-type 5' UTR, 3) a vector control for background.
But depending on the journal, a simple RT-PCR in normal and disease samples can be enough to validate the mutation.
Log in to answer this question.
-
Illumina reads clipped at the expected Cas9 double-strand break site – how to interpret?
written by francois •
-
Job: Postdoctoral Researcher in Bioinformatics and Genetics - Washington, D.C
written by shaolei.teng •
-
Association analysis with SNP
written by cwwong13 •
-
How to validate genes identified computationally
written by MAPK
-
Using Data Tracks in Budding Yeast's Gbrowse
written by abenabba •
-
Sequence Conservation/Multiple alignment
written by Diana
-
Using Rate4Site To Calculate Mutation Rate Per Gene?
written by anin.gregory
-
Stop-Gain Mutation Predicted As Benign By Sift/Polyphen2?
written by michealsmith
-
Convertion Of Mutation Names And Amino Acid Changes To Rs
written by Vova Naumov
-
Getting Conservation Scores For Specific Sites
written by Rubal7
