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ZKO number ZKO1156
Recent progress of this line F2 individuals have been indentified
Gene name C3 (7 of 9)
Ensembl ID C3 (7 of 9) ENSDARG00000090176
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
Cas9 target site 2 sequence
Position in the gene exon4
Restriction enzyme (RE) n/a
PCR primer (forward) 5’- TCTGTTAAACATCATTCGGGA -3’
PCR primer (reverse) 5’- ATGCACATCCATCATCAGCA -3’
Length of expected PCR product (bp) 473
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pSP6-2sNLS-spCas9
Plasmids used for making gRNA pT7-gRNA
Primers used for making gRNA Pr-ZKO
Dosage of injected Cas9 mRNA (ng/ul) 563
Dosage of injected gRNA (ng/ul) 127.6
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1156.pdf
Mutation efficiency in F0 embryos (%) 81.8
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 100% (2/2: Number of positive F0 / Total number of F0 screened)
Wild-type (WT) sequence tggcagctaaGGATTTCGAGAGCGCCGGGCaggtggtgcgctg
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 tggcagctaaGGATTTCGAGAGCGCxxxxxaggtggtgcgctg (-5 bp)
F0-2 tggcagctaaGGATTTCGAGAGCctcatcactcgtttaaGGGCaggtggtgcgctg (+16 bp, -3 bp) tggcagctaaGGATTTCGAGxxxxxxxxGCaggtggtgcgctg (-8 bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 33.3% (1/3: Number of F1 carrier / Total number of F1 screened) tggcagctaaGGATTTCGAGAGCGCxxxxxaggtggtgcgctg (-5 bp)
F0-2 66.7% (2/3: Number of F1 carrier / Total number of F1 screened) tggcagctaaGGATTTCGAGxxxxxxxxGCaggtggtgcgctg (-8 bp) tggcagctaaGGATTTCGAGAGCGCCGxxxxxxtggtgcTctg (-6 bp)
Allele name and predicted product 1156a
tggcagctaaGGATTTCGAGAGCGCxxxxxaggtggtgcgctg; Truncated protein (190 aa)
tggcagctaaGGATTTCGAGxxxxxxxxGCaggtggtgcgctg; Truncated protein (189 aa)
突变创建时间 2014-08-01
填表时间 2013/12/20
修改时间 2016/8/9
PI/负责人 (工作单位)
Genotype Information: