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ZKO number ZKO189
Recent progress of this line F2 generation has been raised
Gene name zgc:136472
Ensembl ID ENSDARG00000058445
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGCGGAGCTTTTAAGAACTGggg
Cas9 target site 2 sequence
Position in the gene exon3
Restriction enzyme (RE) BsrI
PCR primer (forward) ttcacctcactgttccctc
PCR primer (reverse) AATAATCCAAGCACCACGA
Length of expected PCR product (bp) 476
Length of expected RE digestion product (bp) 350+120
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 189
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 digestion30
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO189.pdf
Mutation efficiency in F0 embryos (%) 40
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 20
Wild-type (WT) sequence ATTATTATTGTTTTTGTGTAACTCCTCCCCCAGTTCTTAAAAGCTCCGCCTCCATCCTGACGTGG CTTAAAAGAAGAGCA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 ATTATTATTGTTTTTGTGTAACTCCTCCCCCAGaTTCTTAAAAGCTCCGCCTCCATCCTGACGTGGCTTAAAAGAAGAGCA +1bp
F0-2 ATTATTATTGTTTTTGTGTAACTCCTCCCC---TTCTTAAAAGCTCCGCCTCCATCCTGACGTGGCTTAAAAGAAGAGCA -3bp
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 ATTATTATTGTTTTTGTGTAACTCCTCCCCCAGaTTCTTAAAAGCTCCGCCTCCATCCTGACGTGGCTTAAAAGAAGAGCA +1bp
F0-2 ATTATTATTGTTTTTGTGTAACTCCTCCCC---TTCTTAAAAGCTCCGCCTCCATCCTGACGTGGCTTAAAAGAAGAGCA -3bp
Allele name and predicted product 189a
189b
ATTATTATTGTTTTTGTGTAACTCCTCCCCCAGATTCTTAAAAGCTCCGCCTCCATCCTGACGTG GCTTAAAAGAAGAGCA change splicing
ATTATTATTGTTTTTGTGTAACTCCTCCCCTTCTTAAAAGCTCCGCCTCCATCCTGACGTGGCTTAAAAGAAGAGCA change splicing
突变创建时间
填表时间 2013/12/17
修改时间 2016/12/30
PI/负责人 (工作单位)
Genotype Information: