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ZKO number ZKO76
Recent progress of this line F2 individuals have been indentified
Gene name si:ch211-235f1.1
Ensembl ID ENSDARG00000074381
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GAAGTAATCGCCCTCGACGagg
Cas9 target site 2 sequence
Position in the gene exon 3
Restriction enzyme (RE) Bsl I
PCR primer (forward) gcagtgtaaacaatctggat
PCR primer (reverse) gctgttatcgatataggcat
Length of expected PCR product (bp) 406
Length of expected RE digestion product (bp) 205+201
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 76
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 70
Method to estimate mutation efficiency Restriction enzyme digestionBsl I
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO76.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 30
Wild-type (WT) sequence CAACGGGCCCCAGGGAAGGCGCTGTTTGACCTGGTTTGTTCTCATCTCAACCTCGTCGAGGGCGATTACTTCGGTTTGGAGTTTCAGGATCAGCGGAAAATGATT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 2bp deletion CAACGGGCCCCAGGGAAGGCGCTGTTTGACCTGGTTTGTTCTCATCTCAACCTCGTCGAGGG--ATTACTTCGGTTTGGAGTTTCAGGATCAGCGGAAAATGATT (+0bp, -2bp) 105aa
F0-2 10bp deletion CAACGGGCCCCAGGGAAGGCGCTGTTTGACCTGGTTTGTTCTCATCTCAACCTC------------------ATTACTTCGGTTTGGAGTTTCAGGATCAGCGGAAAATGATT (+0bp, -10bp) 85aa
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 2bp deletion CAACGGGCCCCAGGGAAGGCGCTGTTTGACCTGGTTTGTTCTCATCTCAACCTCGTCGAGGG--ATTACTTCGGTTTGGAGTTTCAGGATCAGCGGAAAATGATT (+0bp, -2bp) 105aa
F0-2 10bp deletion CAACGGGCCCCAGGGAAGGCGCTGTTTGACCTGGTTTGTTCTCATCTCAACCTC------------------ATTACTTCGGTTTGGAGTTTCAGGATCAGCGGAAAATGATT (+0bp, -10bp) 85aa
Allele name and predicted product 76a
76b
TCATCTCAACCTCGTCGAGGG--ATTACTTCGGTTTGGAGTTTCAGGATCAGCGGAAAATGATT 2bp deletion truncated protein 105aa
CAACGGGCCCCAGGGAAGGCGCTGTTTGACCTGGTTTGTTCTCATCTCAACCTC------------------ATTACTTCGGTTTGGAGTTTCAGGATCAGCGGAAAATGATT truncated protein 85aa
突变创建时间
填表时间 2013/12/18
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: