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Phoenix Amphoteric cell line to package lentivirus?

Lentivirus packaging Phoenix Amphoteric Retrovirus

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6 replies to this topic

#1 Epigeneticist

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Posted 17 March 2014 - 12:28 PM

Can the Phoenix Amphoteric cell line be used to package lentivirus or only retrovirus? I cannot find any information online. 

 

Thanks for your help. 


Edited by Epigeneticist, 17 March 2014 - 12:39 PM.


#2 Epigeneticist

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Posted 18 March 2014 - 05:02 PM

FYI: I got the answer straight from the source-

 

The Phoenix Amphoteric cell line can only be used to package retroviruses (no lentivirus).  The packaging proteins from Phoenix do not recognize any of the packaging signals from HIV or other lentis.  Too evolutionarily distant.


#3 Rsm

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Posted 19 March 2014 - 09:09 AM

I regularly use the Phoenix cell line for lentivirus production, works better than HEK293 in my hands. Of course you'll have to provide the necessary vectors in trans...


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#4 Epigeneticist

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Posted 19 March 2014 - 05:51 PM

I regularly use the Phoenix cell line for lentivirus production, works better than HEK293 in my hands. Of course you'll have to provide the necessary vectors in trans...

 

Thanks for the clarification Rsm, Phoenix cells can be used to package lentivirus similar to using 293T cells but only when transfected with the necessary vectors. However, retroviruses can be packaged in Phoenix cells without any additional vectors. 



#5 Epigeneticist

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Posted 19 March 2014 - 05:57 PM

I regularly use the Phoenix cell line for lentivirus production, works better than HEK293 in my hands. Of course you'll have to provide the necessary vectors in trans...

 

I have never considered using Phoenix cells in place of 293T so I am curious. In what regard do they work better? Produce higher titer? More adherent? Less death?



#6 Rsm

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Posted 24 March 2014 - 07:56 AM

Yes, higher titer, usually 3-10 fold. I think it may also depend on how old / what passage number the cells are, maybe our Phoenix are younger than our HEK.

 

If you have problems with cells detaching, I'd recommend adding 0.01% poly-Lysine in PBS to your plates before plating the cells. 5min at RT, remove Lysine and plate the cells straight on, they'll love it. Be warned, you won't be able to detach them even using Trypsin (which is not a real surprise).


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#7 Epigeneticist

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Posted 24 March 2014 - 10:00 AM

Thanks Rsm!







Also tagged with one or more of these keywords: Lentivirus packaging, Phoenix Amphoteric, Retrovirus

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