Strain information
 NBRP Rat No: 0931  Strain name: LE-Pvalbem2(T2A-cre)Koba  Commmon Name: PVA-Cre 2ラット Rat Genome Database
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  F4 (April 2020)
Usage Restrictions  The following specific terms and conditions are requested by the DEPOSITOR. In this event, the recipient of BIOLOGICAL RESOURCE shall obtain a prior written consent on use of it from the DEPOSITOR.
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 脳神経科学一般
 Control Strain  Marker Strain
Gene Affected cre: cre recombinase protein, Pvalb: parvalbumin
Origin Background strain: Long-Evans (Iar:Long-Evans, Institute for Animal Reproduction) Details of the transgene: T2A, nlsCre, and BGHpolyA (about 1.3 kb) are knocked in the fourth exon of the rat Pvalb gene (NCBI Gene ID: 25269). The nls refers to the nuclear localization sequence fused to the N-terminal side of cre. The bGHpolyA refers to the bovine growth hormone poly A signal. Method of production: A complex of Cas9 protein and gRNA was introduced into fertilized rat eggs by electroporation. Combi-CRISPR (Yoshimi K. et al.) was used to induce knock-in. Knock-in rats were selected and crossed with wild-type rats to establish this strain. Sequence features: The intron just before the fourth exon of the Pvalb gene (intron 3) is deleted by NHEJ after double-strand break by one base compared to the wild-type. ---ttggcgggccagaacctcagggg---(wild-type) ---ttggcgggccagaacc-cagggg---(knock-in)
Strain characteristics Knock-in rats expressing cre recombinase under the control of Pvalb gene.
Breeding Conditions Good reproductive performance. Heterozygous and wild-type rats are crossed to maintain the strain.
Genotyping Discriminate by checking for the presence of PCR products at the two sites. (1) Product size: 524 bp 5'-GTGCATTTCCAGAGGTGACAG (upstream of the transgene), 5'-TCGCGAACATCTTCAGGTTCT (cre recombinase) (2) Product size: 705 bp 5'-ACCAGCCAGCTATCAACTCG (cre recombinase), 5'-GGTGGTGTCCGATGGGTACAG (downstream of the transgene) PCR conditions: Quick Taq HS DyeMix (TOYOBO) 96℃ 10sec, 　58℃ 20sec, 68℃ 45sec (30 cycles)
References Combi-CRISPR: combination of NHEJ and HDR provides efficient and precise plasmid-based knock-ins in mice and rats.
Yoshimi K, Oka Y, Miyasaka Y, Kotani Y, Yasumura M, Uno Y, Hattori K, Tanigawa A, Sato M, Oya M, Nakamura K, Matsushita N, Kobayashi K, Mashimo T.
Hum Genet. 2021 Feb;140(2):277-287. doi: 10.1007/s00439-020-02198-4.
Additional strain information