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ZKO number ZKO2
Recent progress of this line F2 generation has been raised
Gene name f7
Ensembl ID ENSDARG00000034862
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAGCTGAAGACGGGGAATCtgg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) T7E1
PCR primer (forward) 5’- gctctcgagtctccattactgc-3’
PCR primer (reverse) 5’- cgtggcctctgtgtgctcgaac -3’
Length of expected PCR product (bp) 275
Length of expected RE digestion product (bp) 195bp+80bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids?used?for?making?gRNA pMD19T-gRNA
Primers?used?for?making?gRNA Pr-ZKO 2
Dosage of injected Cas9 mRNA (ng/ul) 150
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 92
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO2.pdf
Mutation efficiency in F0 embryos (%) 69
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type Indel
Germline transmission efficiency in adult F0 50% (16/32: Number of positive F0 / Total number of F0 screened)
Wild-type (WT) sequence GGAGCTGAAGACGGGGAATCTGG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GGAGCTGAAGACGG-------GG (-7 bp)
F0-2 GGAGCTGAAGAC-------CTGG (-7 bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGAGCTGAAGACGG-------GG (-7 bp)
F0-2
Allele name and predicted product 2a
2b
GGAGCTGAAGACGG-------GG; truncated protein (158 aa)
突变创建时间
填表时间 2013/12/24
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information:
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