C5aR GFP-KI

eagle-i ID

Resource Type

Properties

1. Resource Description

   "A loxP site was inserted upstream of exon 2. A cassette containing (5' to 3') an internal ribosomal entry site (IRES), EGFP, SV40 polyadenlyation sequence, FRT-flanked neo cassette and loxP site was inserted into the 3' UTR." This strain is cryo-preserved. Gene targeting in embryonic stem (ES) cells was performed according to standard protocols. Linearized targeting vector was transfected into ES cells (TL-1, a cell line derived from the 129/SvEv mouse strain). The neomycin and diphtheria toxin genes on the vector allowed for double selection of ES cells that underwent homologous recombination and incorporated GFP and loxP into the <i>C5ar1</i> gene locus on one strand of their genomic DNA. Positive clones were expanded, confirmed in second-round screening, and microinjected into C57BL/6 mouse blastocysts before being transferred to pseudopregnant females. Male chimeras were mated subsequently with female C57BL/6 mice (The Jackson Laboratory) to achieve germline transmission. "GFP is expressed highly on Gr-1⁺CD11b⁺ cells in the blood, spleen and bone marrow, and moderately on CD11b⁺F4/80⁺ circulating leukocytes and elicited peritoneal macrophages. No GFP is detected on resting or activated T lymphocytes, nor on splenic myeloid or plasmacytoid dendritic cells."

2. Additional Name

   C5aR GFP knock-in

3. Related Publication or Documentation

   C5aR expression in a novel GFP reporter gene knockin mouse: implications for the mechanism of action of C5aR signaling in T cell immunity

4. Website(s)

   http://www.informatics.jax.org/javawi2/servlet/WIFetch?page=alleleDetail&key=819124

5. Website(s)

   http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3324670/#!po=37.5000

6. Related Technique

   Transgenic mouse model generation

7. Parental Strain Name

   C57BL/6

8. Biological process studied

   gene expression

9. Genetic Alteration(s)

   C5ar1[tm1Song]

10. Phenotype Findings

    Abnormal neutrophil physiology

11. Phenotype Findings

    Abnormal protein targeting

12. Location

    Wen-Chao Song Laboratory