If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO374
Recent progress of this line F2 individuals have been indentified
Gene name si:dkey-245p14.7
Ensembl ID ENSDARG00000061952
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
Cas9 target site 2 sequence
Position in the gene exon 2
Restriction enzyme (RE) n/a
PCR primer (forward) gcgttgcagtcattctcattg
PCR primer (reverse) gagcaggtccagactacaaact
Length of expected PCR product (bp) 590
Length of expected RE digestion product (bp) 249+341
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 374
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 20
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO374.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 33.3%(2/6)
Wild-type (WT) sequence gcttgggtcatgctgaaacagacgagcagttgcaggatattatatctaagttcctgcctcctgtcc
Germline mutations in selected F0
 (By screening F1 embryos)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 gcttgggtcatgctgaaacagacgagcagtattattatctaagtattatataagtattatatcatattatatctaagttcctgcctcctgtcc (+33 bp, -6 bp)
Allele name and predicted product 374a
gcttgggtcatgctgaaacagacgagcagtattattatctaagtattatataagtattatatcatattatatctaagttcctgcctcctgtcc Truncated protein(36aa)
填表时间 2013/12/18
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: