Hello everyone
I am working on next aim of the project which is to produce mRNA vaccines in plants mitigating the needs to purify them and can be consumed as a part of food. I am planning to target my DNA construct encoding the S gene of Covid19 to the chloroplast genome as they have their own transcription machinery and lack gene silencing. I have few doubts in this regards:
a) I am planning to clone my S gene into deconstructed viral vector construct? shall I perform biolistic method to target my transgene into chloroplast genome? I want stable integration into the chloroplast genome (or is their any other alternative)
b) I will do genotyping to confirm the transgene insertion
c) How I am gonna make sure that S gene are not getting translated and remaining as mRNA for mRNA based vaccine? I can do northern blot but since mRNA gets translated/degraded, how the genetic design will make sure that enough mRNA will be their for therapeutic effect?
d) Once everything is confirmed then offcourse we are going to test in mouse model