My lab recently conducted an RNA sequencing experiment examining differentially expressed genes between a control group and an injury group. At this point, we're just using Illumina BaseSpace to look at our data. Several novel transcripts came up as significantly different, but I'm new to genetics and unsure how to pursue the identity (or importance) of these transcripts.
Hi Meaghan,
Not being an expert on that, few approaches which jumps to mind is:
1. Gene knockdown in mouse (if homologs are there) or appropriate tissue culture cells by ZFN/TALEN/CRISPR. (Llengthy and difficult approach but is most reliable)
2. Gene repression by iRNA or other methods - quick and done mainly in TC and as such less telling, particularly if a gene is developmentally important .
3. Gene overexpression from exogenous constructs - quick and even less likely to give you an interpretable answer, again in TC.
4. Bioinformatics on tissue, organism expression prevalence and sequence variability etc. - even less reliable approach.
Anything else guys?
Use blast2go https://www.blast2go.com/ to identify them and compare them with relevant references.
Cheers
Hi Meaghan,
You can inquire your sequences with BLASTX to identify your sequences, and then understand gene function by reading the reference related to the gene. If you want to further elucidate the gene function, you may overexpress or knockdown the gene. if the gene has mutant, you can complement the mutant to analyze its function. good luck!
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