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ZKO number ZKO78
Recent progress of this line F2 generation has been raised
Gene name zgc:162208
Ensembl ID ENSDARG00000070579
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGCTCTGGAAAGCAGGTCTCtgg
Cas9 target site 2 sequence
Position in the gene exon 2
Restriction enzyme (RE) Bsl 1
PCR primer (forward) CCTCATAAAATGTCTCTCTG
PCR primer (reverse) CATCTTTCCTGTCTTCTCG
Length of expected PCR product (bp) 521
Length of expected RE digestion product (bp) 248+273
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 78
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 30
Method to estimate mutation efficiency Restriction enzyme digestionBsl 1
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO78.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 5/16
Wild-type (WT) sequence TTCTGCTGAATGTGCCCGGCTCTGGAAAGCAGGTCTCTGGTGAGATCTACAGCGTTGAC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GTTATTGCCACCAAACACAACATTCCTTTTCTGCTGAATGTGCCCGGC-----------AGCAGGTCTCTGG 7bp deletion
F0-2 GTTATTGCCACCAAACACAACATTCCTTTTCTGCTGAATGTGCCCAGACTGCTCCAGAATGTGCC----CTGGAAAGCAGGTCTCTGGTGAG 4bp deletion 20bp insertion
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GTTATTGCCACCAAACACAACATTCCTTTTCTGCTGAATGTGCCCGGC-----------AGCAGGTCTCTGG 7bp deletion
F0-2 GTTATTGCCACCAAACACAACATTCCTTTTCTGCTGAATGTGCCCAGACTGCTCCAGAATGTGCC----CTGGAAAGCAGGTCTCTGGTGAG 4bp deletion 20bp insertion
Allele name and predicted product 78a
78b
GTTATTGCCACCAAACACAACATTCCTTTTCTGCTGAATGTGCCCGGC-----------AGCAGGTCTCTGG 7bp deletion Truncated protein 97aa
GACTGCTCCAGAATGTGCC----CTGGAAAGCAGGTCTCTGGTGAG 4bp deletion 20bp insertion truncated protein 72aa
突变创建时间 2014-04-10
填表时间 2013/12/18
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: