How would you use PCR to insert it? What protocol are you using? Genetic modification of animal lines is difficult and takes significant planning.
-- How do you "insert" the DNA with CRISPR? CRISPR doesn't integrate anything, just prompts homologous recombination, which works better with homology, provided by a DNA you provide. How will you get the rat sperm? How will you get transfection quality DNA and then transfect the sperm? What viral vector will you use? Is it biosafety 1? How will you do a polycistronic expression cassette? Given RBS strengths predicted in said organism can you make an educated guess (yes the calculator isn't perfect) on expression levels of each gene required in your target organism given a set of promoters? Do you have the materials do undergo that cloning project?
There is a reason there is yet to be a DIYbio project that uses animals. It is pretty rare to see a successful DIYbio that effectively expresses a multi-gene construct. The highest organism is usually plants. Do you even know if your genes confer to radiation resistance in E coli? If they do, does this function in yeast? Do you have the materials to do this in rats? I don't work with multicellular organisms, but I'm pretty sure this will take some training and some expensive equipment.
-Koeng
On Wednesday, February 3, 2016 at 6:13:10 AM UTC-8, Finn Daffron wrote:
On Wednesday, February 3, 2016 at 6:13:10 AM UTC-8, Finn Daffron wrote:
Thank you for all of your answers. My idea was to use UV lights as radiation. I am glad that people believe that that is a good idea. What do you think of using PCR to insert thermococcus gammatolerans DNA into the sperm of rats as opposed to CRISPR?
On Thursday, January 28, 2016 at 3:11:22 PM UTC-5, Finn Daffron wrote:I am trying to create a batch of thermococcus gammatolerans. This is because I want to use CRISPR to inject thermococcus gammatolerans into rats in order to create radiation resistance with possible implications being people working in nuclear power plants, astronauts, etc. Does anyone have any ideas on how it would be synthesized? Please get back to me.
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