If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1118
Recent progress of this line F1 individuals have been identified
Gene name mb
Ensembl ID ENSDARG00000031952
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGGTGATCTGGCGGGAAGCCcgg
Cas9 target site 2 sequence
Position in the gene exon3
Restriction enzyme (RE) HpaII
PCR primer (forward) 5' CATAATTTGTCTGCGATAC 3'
PCR primer (reverse) 5' TTTACCCTGAAGTTGTTG 3'
Length of expected PCR product (bp) 356
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1118
Dosage of injected Cas9 mRNA (ng/ul) 150
Dosage of injected gRNA (ng/ul) 100
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1118.pdf
Mutation efficiency in F0 embryos (%) 5
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 4.3
Wild-type (WT) sequence GGGAAGCCCGGCGGTGGCGGCTCAT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 CTGGGATTTCTCAGGGTGATCTGGCGGGAGCCCGGCGGTGGCGGCTCATGGAGCGACCG
F0-2 CTGGGATTTCTCAGGGTGATCTGGCGGTGATCTGGCGGTGGCGGCTCATGGAGCGACCG
Allele name and predicted product 1118a
1118b
GGTGATCTGGCGGGA-GCCCGGCGGTGGCGGCTCATtruncated protein 71aa
GGCGGTGATCT-GGCGGTGGCGGCTCATGGAGCGACCGtruncated protein 71aa
突变创建时间 2014-06-30
填表时间 2013/12/17
修改时间 2016/9/28
PI/负责人 (工作单位)
Genotype Information: