If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1003
Recent progress of this line F1 individuals have been identified
Gene name ppp3r1b
Ensembl ID ENSDARG00000069360
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GATGGTGGGGAATAACCTGAagg
Cas9 target site 2 sequence
Position in the gene Exon 5
Restriction enzyme (RE) n/a
PCR primer (forward) GTTAGACAATCCTGAAAGCTG
PCR primer (reverse) GAAGGATATCCTGCCGTCACC
Length of expected PCR product (bp) 441
Length of expected RE digestion product (bp) n/a
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1003
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 40
Survival rate of F0 embryo @ 1dpf (%) 28
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1003.pdf
Mutation efficiency in F0 embryos (%) 30
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 40% (2/5)
Wild-type (WT) sequence GGTGCTGAAGATGATGGTGGGGAATAACCTGAAGGACACTCA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGTGCTGAAGATGATGGTGGGGAATAACACTCAACTGGAAGGACACTCA (-2,+9bp)
F0-2
Allele name and predicted product 1003a
1003b
GGTGCTGAAGATGATGGTGGGGAATAACACTCAACTGGAAGGACACTCA protein (146aa)
突变创建时间
填表时间 2014/1/2
修改时间 2016/11/11
PI/负责人 (工作单位)
Genotype Information: