Long-term GFP expression without selection

Gerstein, Rachel Rachel.Gerstein at umassmed.edu
Wed Mar 27 14:36:54 EST 2002


Hi

are you using mouse or human cells ? there is a good episomal systems for
human cells:

Grignani F, Kinsella T, Mencarelli A, Valtieri M, Riganelli D, Grignani F,
Lanfrancone L, Peschle C, Nolan GP, Pelicci PG.High-efficiency gene transfer and
selection of human hematopoietic progenitor
cells with a hybrid EBV/retroviral vector expressingthe green fluorescence
protein. Cancer Res. 1998 Jan 1;58(1):14-9.
Kinsella TM, Nolan GP.Episomal vectors rapidly and stably produce high-titer recombinant
retrovirus.Hum Gene Ther. 1996 Aug 1;7(12):1405-13.

or you could use a retroviral vector.

what is the problem for your system  with making stable integrated lines
using drug selection ?

Good luck
Rachel

=======================================================
Rachel M. Gerstein, Ph.D.
Department of Molecular Genetics and Microbiology
Graduate Program in Immunology/Virology
University of Massachusetts Medical School
55 Lake Avenue North
Worcester, MA 01655-0002
(508) 856-1044
(508) 856-5920 (FAX)


> ----------
> From:		jack
> Sent:		Monday, March 25, 2002 1:20 PM
> To:	Cytometry Mailing List
> Subject:	Long-term GFP expression without selection
>
> Dear Flowers:
>
> Can anyone suggest a GFP vector which can stay stably in dividing cells
> for a long time (such as 1 month) without G418 or other selection? We want
> to try an in vivo study and need the information about the dynamic change
> of targeted cell number. The GFP vector we tried is with CMV promoter and
> the percentage of GFP(+) cells (PC-3) goes down very fast when cultured in
> dishes without selection (even we cultured them from a single cell). Any
> suggestion is deeply appreciated.
>
> Best Wishes,
>
> Jack
>
>
>



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