If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO169
Recent progress of this line F1 individuals have been identified
Gene name grin2ab
Ensembl ID ENSDARG00000070543
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAGGAAGCCCGTTCGCTAggg
Cas9 target site 2 sequence
Position in the gene exon2
Restriction enzyme (RE) T7
PCR primer (forward) tgggaccttcagagcgtagtgac
PCR primer (reverse) catcaggtatccggccttcttct
Length of expected PCR product (bp) 358
Length of expected RE digestion product (bp) 148+210
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 169
Dosage of injected Cas9 mRNA (ng/ul) 200
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 70
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO169.pdf
Mutation efficiency in F0 embryos (%) 20
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0 15
Wild-type (WT) sequence cccgaggaccgagctcatgtgcagctcaaacgcatccagtcgcccgtcatcctcctctactgttccaaagatgaagcagcgctggttatggaggaagcccgttcgctagggctcacaggggctggctttgtgtggattgttcctagcctgacgacagggaatctcgaacatatcccggaagcctttccggcagggatgat
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GCAGCGCTGGTTATGGAGGAAGCCCGTTCGGGCTCACAGGGGCTGGCTTTGTGTGGATTG (-4bp)
F0-2 cccgaggaccgagctcatgtgcagctcaaacgcatccagtcgcccgtcatcctcctctactgttccaaagatgaagcagcgctggttatggagggctcacaggggctggctttgtgtggattgttcctagcctgacgacagggaatctcgaacatatcccggaagcctttccggcagggatgat(-16bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GCAGCGCTGGTTATGGAGGAAGCCCGTTCGGGCTCACAGGGGCTGGCTTTGTGTGGATTG (-4bp)
F0-2 cccgaggaccgagctcatgtgcagctcaaacgcatccagtcgcccgtcatcctcctctactgttccaaagatgaagcagcgctggttatggagggctcacaggggctggctttgtgtggattgttcctagcctgacgacagggaatctcgaacatatcccggaagcctttccggcagggatgat(-16bp)
Allele name and predicted product 169a
169b
truncated protein (248aa)
truncated protein (244aa)
突变创建时间 2014-09-05
填表时间 2014/3/19
修改时间 2016/9/23
PI/负责人 (工作单位)
Genotype Information: