If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO981
Recent progress of this line F2 individuals have been indentified
Gene name herc3
Ensembl ID ENSDARG00000075887
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGTCTGGGAACAGCTGAAGagg
Cas9 target site 2 sequence
Position in the gene Exon 3
Restriction enzyme (RE) Msp A1l(CMG|CKG)
PCR primer (forward) TATGTTTTCGCAATACTCAAC
PCR primer (reverse) CTTACTCCGAATGGCAGA
Length of expected PCR product (bp) 469
Length of expected RE digestion product (bp) 228+241
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 981
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 85
Method to estimate mutation efficiency Restriction enzyme digestionRE digestion (Msp A1l)
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO981.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 18%(2/11)
Wild-type (WT) sequence GGTCTGGGAACAGCTGAAG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGTCTGGGAACAGCTACAGGAAG(+4bp)
F0-2
Allele name and predicted product 981a
981b
GGTCTGGGAACAGCTACAGGAAG; Truncated protein(132aa)
突变创建时间 2014-03-13
填表时间 2013/12/18
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information: