STOCK Tg(Pgk1-Ccnb1/EGFP)1Aklo/J

Stock number: 023345
Product name: CycB1-GFP
Research areas: Cell Biology Research, Research Tools

Description

CycB1-GFP reporter mice express GFP during S-G2-M stages of the cell cycle.

Detailed description

CycB1-GFP transgenic mice express enhanced green fluorescent protein (EGFP) fused to the 105 N-terminal residues of the Cyclin B1 (Ccnb1) gene. This portion of Cyclin B1 (known as the "destruction box") is ubiquitinated by APC/C complex, directing the protein for degradation when APC/C is active. Due to the cell cycle stage specific nature of APC/C activity, only cells that are in S-G2-M stages of the cell cycle express the CycB1-GFP protein. Expression is driven by the housekeeping, constitutive mouse phosphoglycerate kinase 1 (Pgk1) promoter and the reporter is detected in a range of tissues in vivo. The transgene was introduced using a viral vector.

Of note, the transgene was introduced using a viral vector and is integrated in multiple loci. These mice are kept mostly inbred but with time, variation in the level of expression is evident. This becomes more frequent when mice are outbred. As such, genotyping the mice by PCR does not always reflect the transgene expression level. Monitoring GFP level in ear fibroblasts prepared from individual mice is the preferred method for genotyping. Mice with good expression are chosen for analysis and for further breeding.

Development

The CycB1-GFP reporter mouse line was generated via lentitransgenesis; a self-inactivating lentiviral vector was designed with 105 N-terminal residues of the Cyclin B1 (Ccnb1) gene and an enhanced green fluorescence protein (EGFP) sequence under control of a constitutive mouse phosphoglycerate kinase 1 (Pgk1) promoter. CycB1-GFP-expressing lentivirus were microinjected into the perivitelline space of one-cell embryos. Morula and blastocyst stage embryos were implanted into pseudopregnant recipient mice. Founder males, with multiple insertion sites, were bred to females to establish the CycB1-GFP lentitransgenic mouse line. These mice were maintained on a mixed background. Upon arrival at The Jackson Laboratory Repository, mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony.

Allele

Symbol: Tg(Pgk1-Ccnb1/EGFP)1Aklo
Name: transgene insertion 1, Agnes Klochendler
MGI accession ID: MGI:5515374
Generation method: Transgenic
Attributes: Inserted expressed sequence
Synonyms: CycB1-GFP
Marker symbol: Tg(Pgk1-Ccnb1/EGFP)1Aklo
Marker name: transgene insertion 1, Agnes Klochendler
Chromosome: UN
Strain of origin: Not Specified
Expressed genes: Ccnb1,cyclin B1,mouse, laboratory; GFP,Green Fluorescent Protein,
Site of expression: Green fluorescent protein is expressed in a range of tissues during S-G2-M stages of the cell cycle.
Molecular note: The transgene was generated via lentitransgenesis; a self-inactivating lentiviral vector was designed with 105 N-terminal residues of the Cyclin B1 (Ccnb1) gene (harboring the APC/C ubiquitination site responsible for its specific degradation in G0/G1 cells) and an enhanced green fluorescence protein (EGFP) sequence under control of a constitutive mouse phosphoglycerate kinase 1 (Pgk1) promoter. Line 1 was generated.