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ZKO number ZKO190
Recent progress of this line F2 generation has been raised
Gene name ubn1
Ensembl ID ENSDARG00000063169
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GAATATGGGCTTGCTAAGAagg
Cas9 target site 2 sequence
Position in the gene exon5
Restriction enzyme (RE) DdeI
PCR primer (forward) GAAGGCTTTCACAGTTATT
PCR primer (reverse) TCAGTTGAATATCGGAGAC
Length of expected PCR product (bp) 486
Length of expected RE digestion product (bp) 289+197
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 190
Dosage of injected Cas9 mRNA (ng/ul) 500
Dosage of injected gRNA (ng/ul) 100
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency Restriction enzyme digestionDdeI digest
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO190.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 2/12
Wild-type (WT) sequence GACAAGAAGATAATGAAAAAGAAAAGAGATGAATATGGGCTTGCTAAGAAGGATGAAGAGGAATTCACAAAGAGgttaa
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GACAAGAAGATAATGAAAAAGAAAAGAGATGAATATGGGCTTGCTAttcAGgatggatgaagaggaAAGGATGAAGAGGAATTCACAAAGAGgttaa -2bp,+20bp premuture stop codon TGA appears
F0-2 GACAAGAAGATAATGAAAAAGAAAAGAGATGAATATGGGCTTGCTA-GAAGGATGAAGAGGAATTCACAAAGAGgttaa -1bp
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GACAAGAAGATAATGAAAAAGAAAAGAGATGAATATGGGCTTGCTAttcAGgatggatgaagaggaAAGGATGAAGAGGAATTCACAAAGAGgttaa -2bp,+20bp premuture stop codon TGA appears
F0-2
Allele name and predicted product 190a
190b
GAATATGGGCTTGCTAttcAGgatggatgaagaggaAAGGAT truncated protein(90aa)
突变创建时间 2015-07-15
填表时间 2013/12/17
修改时间 2016/12/30
PI/负责人 (工作单位)
Genotype Information: