Basic Information
Strain Name
SD-Gt (ROSA)26Sor tm1(CAG-LSL-3XGFP) /Bcgen
Stock Number
B-CR-017
Common Name
B-3XGFP cKI rats
Source/Investigator
Bcgen (Beijing Biocytogen Co., Ltd)
Related Genes
GFP fluorescent protein
Species
Sprague Dawley
Appearance
White
Genotypes
GFP cKI (wt/mut)
Development
A designed targeting sequence containing pCAG-loxP-3*STOP-loxP- 3*GFP-WPRE-bGHpA was inserted to the Rosa26 between exon 1 and exon 2 using CRISPR/Cas9 technology. This strain was maintained on a Sprague Dawley genetic background.
Description
These GFP cK I rats knockin rats have a floxed-STOP cassette preventing expression of the downstream bicistronic sequence (3*GFP). Although under control of a CAG promoter, widespread expression of GFP is prevented by the STOP cassette. After exposure to Cre recombinase, expression of GFP will be observed.
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Targeting Strategy
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Details
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Genotyping Information
Primer Sequence (5’-3’) Tm (℃) Product size
(bp)GFP cKI primer 1 TTGTATTGGAGACAAGAAGCACTTGCTC 60 WT: 646
Mut: 6583GFP cKI primer 2 TTGATAGGGCTGCAGAAGTGGGAG 61 GFP cKI primer 3 TGCACTGTGTTTGCTGACGC 60 Mut: 230 GFP cKI primer 4 ATCCAGGTGGCAACACAGGC 63 Polymerase: Kod-FX
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References
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1. Ma Y, Zhang X, Shen B, Lu Y, Chen W, Ma J, Bai L, Huang X, Zhang L. 2014. Generating
rats with conditional alleles using CRISPR/Cas9. Cell Res. Jan;24(1):122-5. [PubMed: 24296780]2. Ma Y1, Ma J, Zhang X, Chen W, Yu L, Lu Y, Bai L, Shen B, Huang X, Zhang L. 2014.
Generation of eGFP and Cre knockin rats by CRISPR/Cas9. FEBS J. Sep;281(17):3779-90.
[PubMed: 24296780]
