ZKO number ZKO1
Recent progress of this line F2 generation has been raised
Gene name zgc:163025
Ensembl ID ENSDARG00000076045
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGGGACGGTGAAGATCTCTCtgg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) T7E1
PCR primer (forward) 5’-gtgtccatgatctgcagtgttc-3’
PCR primer (reverse) 5’-gtgttctatattcttgtttaatg -3’
Length of expected PCR product (bp) 218
Length of expected RE digestion product (bp) 158bp+60bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1
Dosage of injected Cas9 mRNA (ng/ul) 150
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 62.5
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1.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 66.7% (m/n: Number of positive F0 / Total number of F0 screened) (2/3)
Wild-type (WT) sequence GGGGACGGTGAAGATCTCTCTGG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GGGGACGGTGAAGATAT----GG ( 5bp deletion +1bp insertion)
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGGGACGGTGAAGATAT----GG ( 5bp deletion + 1bp insertion)
F0-2
Allele name and predicted product 1a
1b
GGGGACGGTGAAGATAT----GG; truncated protein (195 aa)
突变创建时间 2014-07-15
填表时间 2013/12/24
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotye Information: