do all the lentivectors have the same insert size limit?

Aawara Chowdhury aawara at FEMA-trailer.org
Thu Sep 14 20:47:41 EST 2006


In <1158248120.311491.319740 at k70g2000cwa.googlegroups.com>,
 dnhkng at googlemail.com <dnhkng at googlemail.com> wrote:

> so, i read the 2001 paper on lentivirus insert size, and it seems there
> is a logarithmic decrease in expression level on increasing insert
> length... damn...
>
> but that was a 2001 paper, so maybe things have changed a bit.
>
> I want to use a 2kb neuron specific promoter with my 5kb construct.
> thats quite a bit past invitrogens virapower 5.5kb limit recomendation.
> also, i would love to be able to add an IRES GFP or even a H2B-GFP
> histone binding fluorescent protein as a marker. that would take me to
> around 10kb.
>
> my understanding is that the wildtype lentivirus has a 10kb genome, but
> lentivectors require about 4.5kb, leaving about 5.5kb for the insert.
>
> does anyone know if the lentivectors out there are identical with
> regards to insert length, or if maybe one has a reduced set of wildtype
> DNA so the vector can accomodate a bigger insert?
>
> thanks in advance for your help, advice or anything you can add 
> david
>

The largest insert sizes that I've put into either a retroviral vector
or a lentiviral vector is about 6 kb.  The efficiency of packaging RNAs
longer than 8 kb - 10 kb is very low.

The exception are ASLV-based vectors, which can have a total size of
upto 14 kb.  There's a paper from Steve Goff's lab that is about 6 - 7 
years old where he demonstrated that like lentiviruses, avian retroviruses
can also infect non-dividing cells.  You might want to consider using
one of Steve Hughes ASLV vectors.

AC
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