Gene targeting in mice has been accomplished traditionally with embryonic stem cells derived from 129 strains.  Because of their genetic variability, poor reproductive performance and abnormal behaviors, the 129 knockout mice are therefore preferably backcrossed to C57BL/6 mice.  C57BL/6 is a reference strain of the mouse genome, many mutants are maintained on this background and widely used in genetic, immunological and behavioral studies, which provide significant advantages of producing knockout mice directly in the C57BL/6 strain.  Although the first C57BL/6 embryonic stem cell line was derived as early as 1991, a few lines have been successfully used in the past decade.

GTTF has derived multiple ES cell lines from the following mouse strains: C57BL/6, C57BL/6J-Tyr^c-2J and NZM2328.  The C57BL/6 and C57BL/6J-Tyr^c-2J ES cells lines can be used for gene targeting directly onto the C57BL/6 background, therefore circumventing the need for backcrossing.  NZM2328 is a congenic strain and a valuable mouse model for systemic lupus erythematosus (SLE) developed at UVa.  The ES cells from NZM2328 will facilitate direct exploration of SLE susceptible genes in this model strain.

GTTF also has stocks of popular germline competent 129 ES cell lines R1, CJ7, W4, RW4 and C57BL/6 ES cell lines.  Appropriate MTAs are required before we can fulfill the requests for these ES cell lines.

Mouse ES cell lines used in GTTF
129 strains - New nomenclature
R1 (Progeny of 129X1/SvJ and 129S1/Sv-+p +Tyr-c MgfSl-J/+) - Nagy A et al, Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8424-8
CJ7 (129S3/SvImJ-+p +Tyr-c +Mgf-SlJ/J) - Swiateck PJ and Gridley T, Genes Dev. 1993 Nov;7(11):2071-84
- matching isogenic BAC library CT7 from California Institute of Technology and sold by Invitrogen
W4 (129S6/SVEvTac) - www.taconic.com/emerging/ESCells/ES_WEB.htm
- matching isogenic Lambda library from Stratagene (now Agilent) or matching isogenic BAC library RP22 from Roswell Park Cancer Institute and sold by Invitrogen
RW4 (129X1/SvJ) - Heusel JW et al, Cell 1994 Mar 25;76(6):977-87
- matching isogenic Lambda library from Stratagene (now Agilent)
4A4 (C57BL/6, GTTF-derived) - Reference available from GTTF.
B1 (C57BL/6J-Tyrc-2J, GTTF-derived) - Reference available from GTTF.
Bruce4 (Thy1.1 congenic C57BL/6 strain) - Kontgen F et al, Int Immunol. 1993 Aug;5(8):957-64.
#693 (C57BL/6) - stemcells.atcc.org/cellp.cfm
- matching isogenic BAC library RP23 from Roswell Park Cancer Institute and sold by Invitrogen

Mouse ES cell lines available from commercial sources
Stem Cell Center - stemcells.atcc.org

ES cell karyotyping
Applied Genetics Laboratories - www.appliedgenetics.com
Chrombios GmbH - www.chrombios.com

Mouse primary embryonic fibroblast (PMEF)
ATCC - stemcells.atcc.org/cellp.cfm