Hello folks,
I have read this somewhere:
"Design primers in the conserved fragments if family genes need to be silenced. However If specific genes need to be silenced, non-conservative regions or 3'non-coding regions can be selected for primer designing. The length of insertion fragment is 150-1000bp, where silencing effect is better in 300-800bp fragment. Too small will reduce the silencing efficiency. Too large will be unstable to cause fragment deletion. Silencing the non-conservative region at the 3'end of the target gene of a single gene selection ensures the silencing effect and specificity."
If someone has experienced it, please let me know..
Thanks
Lee, et al. (2015). Virus induced gene silencing (VIGS) for functional analysis of wheat genes involved in Zymoseptoria tritici susceptibility and resistance. Fungal genetics and biology. doi:10.1016/j.fgb.2015.04.006
"When possible, select at least two preferably non-overlapping regions of the gene of interest for VIGS analyses. Observation of the same phenotype induced by silencing using each of the two or more independent VIGS constructs is a good indication that the phenotype is due to specific silencing of the intended target gene, therefore allowing greater confidence in the obtained results.
When attempting to silence an individual member of a gene family consider selecting the sequences from the 3′- or 5′-UTR regions, which are generally more variable than the CDS. This should minimize the risk of off-target silencing. On the other hand, in cases when a great deal of functional redundancy is expected among different gene family members, it should be possible to design VIGS construct(s) from the conserved gene regions in order to target several or even all gene family members simultaneously."
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