Strain information
NBRP Rat No: 1046
Strain name
: LE-
Adora2a
em2(flpo)Koba
Commmon Name: A2AR-Flpo-2
Principal Investigator:
Kazuto Kobayashi Fukushima Medical University 1 Hikarigaoka Fukushima-shi 960-1295 Fukushima Japan
Tel: 024-547-1667 Fax: 024-548-3936
Email: kazuto@fmu.ac.jp
Preservation Status:
Embryo
Sperm
Living Animals
Coat Color
白黒
Inbred Generations
F9 (Nov. 2025)
Usage Restrictions
The user requires a collaboration with the depositor. Consult with the depositor about details beforehand.
Genetic Status
Inbred
Segregating
Congenic
Consomic
Recombinant
Coisogenic
Spont. Mutant
Transgene
Ind. Mutant
Category Other
Comercial Availability
Research Category
Diabetes Obesity
Neurobiology
Ophthalmology
Dentistry
Cardio Hypertension
Cancer
Metabolism
Otorhinology
Immunology
Infectious
Osteosis
Internal Organ
Dermatology
Reproduction
Development
Behavior
Hematology
Urology
Pharmacology
Research Area Others
Control Strain
Marker Strain
Gene Affected
Adora2a (adenosine A2a receptor)
Origin
Background strain: Long-Evans (Iar:Long-Evans, Institute for Animal Reproduction). Using the CRISPR/Cas9 system (Combi-CRISPR), a double-strand break was targeted to the sequence near the stop codon in the third exon of the adenosine A2a receptor (Adora2a) gene locus in fertilized rat eggs, and a modified Flp recombinase gene (Flpo) was knocked in via homologous recombination. Therefore, Flpo expression is synchronized with the spatiotemporal regulation of Adora2a expression. In the A2AR-Flpo-2 line, a 40-base insertion (underline) is observed in the 3'UTR of the Adora2a gene due to NHEJ after a double-strand break induced by Combi-CRISPR. Target site sequence: TGCTTCTGCTTCGTGCTACG(GGG) A2AR-Flpo-2 knock-in line: TGCTTCTGCTTCGTGCT
CCCACTTTGAACTACAAAGAAGTTCCCACTTTGAACTACT
ACG(GGG) ( ) indicates the PAM sequence.
Strain characteristics
Flpo is expressed in accordance with the spatiotemporal regulation of Adora2a expression.
Breeding Conditions
Good breeding performance. Maintained by mating heterozygous rats with Long-Evans rats.
Genotyping
Forward primer: 5'-CACTGAGGGAGAGCAGGTTAGC (within exon 3) Reverse primer: 5'-AGCTCCTATATTCTAGCATCGG (within 3'UTR) Amplified fragment size: Wild-type: 591 bp; Knock-in: 1990 bp In heterozygotes, both wild-type and knock-in amplifications are detected simultaneously. An annealing temperature of 60℃ (or higher) is recommended.
References
準備中
Additional strain information