B6;FVB-Tg(Cx3cl1-EGFP/Rpl10)ES26Htz/J

Stock number: 037194
Product name: Cx3cl1-bacTRAP
Research areas: Cell Biology Research, Neurobiology Research, Research Tools

Description

Cx3cl1-bacTRAP transgenic mice express EGFP-tagged ribosomal protein L10a (Rpl10a) from the mouse Cx3cl1 (chemokine (C-X3-C motif) ligand 1) promoter/regulatory elements. The mutation enables affinity purification of polyribosomes for analysis of ribosome‐bound mRNAs using the translating ribosome affinity purification (TRAP) method. This strain will be most useful to investigators examining molecular properties of dentate gyrus granule cells.

Detailed description

Cx3cl1-bacTRAP transgenic mice express EGFP-tagged ribosomal protein L10a (Rpl10a) from the mouse Cx3cl1 (chemokine (C-X3-C motif) ligand 1) promoter/regulatory elements. EGFP/Rpl10 is expressed in distinct populations of neurons throughout the brain, including excitatory neurons in the neocortex and granule cells in the dentate gyrus. The mutation enables affinity purification of polyribosomes for analysis of ribosome‐bound mRNAs using the translating ribosome affinity purification (TRAP) method.

The expression pattern of the bacTRAP is more restricted than that described in the Allen Brain Atlas, but aligns very well with that of the GENSAT EGFP reporter. The fluorescence of EGFP can be directly detected in cortex layer 5 and the dentate gyrus. Immunocytochemistry is necessary for other regions, including cortex layers 2/3 and 6. Images from this mouse line can be viewed on the GENSAT (Gene Expression Nervous System Atlas) website.

The integration site and copy number of the BAC transgene are unknown. Hemizygous mice are viable and do not display any obvious physical or behavioral abnormalities. The viability/fertility of homozygotes has not been assessed, and the donating laboratory does not recommend generating them due to the potential for rearrangement of the BAC concatemers and expression pattern changes.

Development

EGFP reporter fused to Rpl10a (ribosomal protein L10a) was introduced immediately upstream of the ATG start site of the mouse Cx3cl1 gene on BAC RP23-238O24. The BAC also carries sequences for the following genes, but their expression has not been confirmed: Ccl22, Ciapin1, Ccl17, Dok4, Coq9, Ccdc102a, and Adgrg5. The modified BAC was microinjected into the pronucleus of FVB/N zygotes and randomly integrated into the genome of the mouse. Resultant founder mice were crossed with C57BL/6J animals for one generation. The integration site and copy number of the BAC transgene are unknown. This strain was maintained on mixed C57BL/6J and FVB genetic background by the donating laboratory. This is founder line ES26.

Upon arrival at The Jackson Laboratory, sperm was cryopreserved. Cryorecovery/live colony development will involve C57BL/6J (Stock No. 000664). (April, 2022)

Allele

Symbol: Tg(Cx3cl1-EGFP/Rpl10)ES26Htz
Name: transgene insertion ES26, Nathaniel Heintz
MGI accession ID: MGI:7260284
Generation method: Transgenic
Attributes: Reporter
Synonyms: Cx3cl1-bacTRAP
Marker symbol: Tg(Cx3cl1-EGFP/Rpl10)ES26Htz
Marker name: transgene insertion ES26, Nathaniel Heintz
Chromosome: UN
Strain of origin: FVB/N
Molecular note: EGFP reporter fused to Rpl10a (ribosomal protein L10a) is introduced immediately upstream of the ATG start site of the mouse Cx3cl1 gene on BAC RP23-238O24. The BAC also carries sequences for the following genes, but their expression has not been confirmed: Ccl22, Ciapin1, Ccl17, Dok4, Coq9, Ccdc102a, and Adgrg5.