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ZKO number ZKO92
Recent progress of this line F2 individuals have been indentified
Gene name spry2
Ensembl ID ENSDARG00000078308
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GTACACGGAGGGCCCGAcgg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) Apa I (GGGCC^C)
PCR primer (forward) CTCATAGGACTTCAGATGTGT
PCR primer (reverse) GGTGGTACTGTGTGAACCAT
Length of expected PCR product (bp) 430
Length of expected RE digestion product (bp) 219+211
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 92
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%)
Method to estimate mutation efficiency Restriction enzyme digestionApa 1
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO92.pdf
Mutation efficiency in F0 embryos (%) 20
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 20
Wild-type (WT) sequence TGGCAGGCCACAGACTGGGGACTCTGAGCCTTGGGATGGTCAGGTGCTCTCCCTGGATCAGATCAGGACCATCCGTGGTAGCAATGAGTACACGGAGGGCCCGACGGTGGCGCCACGGCCCCCGGCCTCTGAGCAAAAAACGGATTCACAAGCTTCCAGCCCAACTTCCACCAGCTCTATTGAGCT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 ZKO92a 1bp insertion 20bp deletion TGGCAGGCCACAGACTGGGGACTCTGAGCCTTGGGATGGTCAGGTGCTCTCCCTGGATCAGATCAGGACCATCCGTGGTAGCAATGAGTACG-------------------------------------GCCACGGCCCCCGGCCTCTGAGCAAAAAACGGATTCACAAGCTTCCAGCCCAACTTCCACCAGCTCTATTGAGCT (+1bp, -20 bp)
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 ZKO92a 1bp insertion 20bp deletion TGGCAGGCCACAGACTGGGGACTCTGAGCCTTGGGATGGTCAGGTGCTCTCCCTGGATCAGATCAGGACCATCCGTGGTAGCAATGAGTACG-------------------------------------GCCACGGCCCCCGGCCTCTGAGCAAAAAACGGATTCACAAGCTTCCAGCCCAACTTCCACCAGCTCTATTGAGCT (+1bp, -20 bp)
F0-2
Allele name and predicted product 92a
92b
GCAATGAGTACG-------------------------------------GCCAC truncated protein(84aa)
突变创建时间
填表时间 2013/12/18
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: