What is the biggest insert for a viral vector? - (Feb/06/2009 )
scolix on Feb 6 2009, 05:22 PM said:
Kami23 on Feb 6 2009, 06:12 PM said:
the only problem with number 4 is our gene doesnt exist in mouse (well it did, but has now lost all function). Ill try to find a way to use these ideas in my project planning I will be back with more questions though 
AAV serotype 2 is good for retinal ganglion cells. But ofcourse has a limitation. There is one paper which cut a gene into 2 and cloned each fragment into 2 different AAV and injected the virus. They later detected the complete protein. Just one way of using existing vectors for experiments
you dont happen to have the citation for that paper do you? im going to plan everything thoroughly this weekend so i can present my plan to my supervisor on monday. I just thought of something but now i cant remeber.... ah well it will come back to me lol
I remeber now... say I used the cDNA then would I have to get a vector containing the promotor or would i have to somehow make my own?
Kami23 on Feb 6 2009, 06:25 PM said:
scolix on Feb 6 2009, 05:22 PM said:
Kami23 on Feb 6 2009, 06:12 PM said:
the only problem with number 4 is our gene doesnt exist in mouse (well it did, but has now lost all function). Ill try to find a way to use these ideas in my project planning I will be back with more questions though AAV serotype 2 is good for retinal ganglion cells. But ofcourse has a limitation. There is one paper which cut a gene into 2 and cloned each fragment into 2 different AAV and injected the virus. They later detected the complete protein. Just one way of using existing vectors for experiments
you dont happen to have the citation for that paper do you? im going to plan everything thoroughly this weekend so i can present my plan to my supervisor on monday. I just thought of something but now i cant remeber.... ah well it will come back to me lol
I think its from a Xiao lab. Try searching for it in pubmed. I will try to search and if I find it will post it here.
Kami23 on Feb 6 2009, 09:04 AM said:
what would you say is the biggest insert you could put into a viral vector and then transfect into a mouse?
Did you get your answers? I am planning to subclone 7.5kb cDNA in retrovirus or lentivirus vector so that I can make stable cell pools.
This may be a little late, but there are several publications out there using IN VIVO and EX VIVO electroporation in the eye - retina and otherwise. I am told it can be very selective spacially once you become proficient at the technique.
Make sure you get the cDNA from the same tissue you plan to express it in under similar conditions. mRNA can be spliced very differently and this can be tissue specific and cell activation can cause the same effect. So your cDNA might look very different and have actually different properties than the one that is original for the tissue ...