The L3mbtl2 floxed exons 9-14 allele has loxP sites flanking the sequences encoding its three C-terminal malignant brain tumor (MBT) domains. Exposure to Cre recombinase creates a null allele. These mice may be useful in studying early embryonic development, methylated histone-binding and the polycomb repressive complex (e.g., PRC1.6).
Hanno R Hock, Massachusetts General Hospital
Malignant brain tumor (MBT) domain chromatin reader proteins bind to methylated histone lysine residues and associate with chromatin remodeling complexes. One such protein, L3MBTL2, functions in early embryonic development and methylated histone-binding, and is an essential component of an atypical Polycomb complex (termed polycomb repressive complex 1.6 or PRC1.6).
The L3mbtl2 floxed exons 9-14 allele has loxP sites flanking the sequences encoding its three C-terminal MBT domains. Mice homozygous for the L3mbtl2 floxed allele are viable and fertile with no reported gross phenotypic or behavioral abnormalities.
Upon exposure to Cre recombinase, the floxed sequences are deleted - resulting in a null allele (no mRNA or protein is expressed).
For example, when bred to germline Cre-expressing mice (e.g., GATA1-Cre), the resulting L3mbtl2 global knockout homozygotes die in utero with failure of gastrulation. Mice heterozygous for the L3mbtl2 global knockout were viable and fertile with no reported abnormalities.
The L3mbtl2 floxed exons 9-14 allele was created by Dr. Hanno Hock (Massachusetts General Hospital). First, a targeting vector was designed to insert a frt::neoR::frt::loxP upstream of exon 9, and a loxP site downstream of exon 14 of the L3mbtl2 gene on chromosome 15. The targeting vector was electroporated into (C57BL/6 x 129S4/SvJae)F1-derived V6.5 embryonic stem (ES) cells. ES cells with the "L3mbtl2 FL+N (Floxed-Neo+)" genotype were transiently transfected with a FLP-expressing plasmid to remove the neoR cassette. The resulting ES cells with the "L3mbtl2 FLΔN (FloxedΔNeo)" genotype (frt::loxP::exons 9-14:loxP) were identified and injected into blastocysts. Chimeric animals were mated with C57BL/6J mice for germline transmission - establishing the L3mbtl2 floxed mice on a C57BL/6;129S4 background. The donating investigator reports the L3mbtl2 floxed colony was maintained on a C57BL/6;129S4 background by breeding L3mbtl2 floxed mice together for several generations. At some point, the mice were also bred to other mutant mice (containing neo; unknown genetic background). In 2017, L3mbtl2 floxed males with black coat color were sent to The Jackson Laboratory Repository. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to remove the neo.
|Allele Name||targeted mutation 1.1, Hanno Hock|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||L3mbtl2, L3MBTL2 polycomb repressive complex 1 subunit|
|Strain of Origin||(C57BL/6 x 129S4/SvJae)F1|
|Molecular Note||LoxP sites flanking exons 9-14 and a neomycin gene flanked by FRT sites were inserted via homologous recombination. Flp-mediated recombination removed the neomycin gene.|
Mice homozygous for the L3mbtl2 floxed allele are viable and fertile with no reported gross phenotypic or behavioral abnormalities. When maintaining a live colony, heterozygous mice may be bred together or to wildtype mice from the colony. Alternatively, homozygous mice may be bred together.
When using the L3mbtl2 floxed exons 9-14 mouse strain in a publication, please cite the originating article(s) and include JAX stock #029979 in your Materials and Methods section.
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