If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1020
Recent progress of this line F1 progeny has been raised
Gene name asna1
Ensembl ID ENSDARG00000018190
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAGGCCATGAGTGCTTTCCCtgg
Cas9 target site 2 sequence
Position in the gene exon3
Restriction enzyme (RE) CC/TGG(Bst NI)
PCR primer (forward) TGTCTGTCTGCAGGAGATCG
PCR primer (reverse) CTTGAACGCGAGTCTGACCT
Length of expected PCR product (bp) 180
Length of expected RE digestion product (bp) 123+57
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1020
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency Restriction enzyme digestionenzyme
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1020.pdf
Mutation efficiency in F0 embryos (%) 94
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 66.6
Wild-type (WT) sequence GGAGGCCATGAGTGCTTTCCCTGGCATCGATGAGGCCATGAGCTACGCAGAGGTCA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 ATGGGGAAAAAGATGATGCAGGAGGCCATGGCCATGAGCTACGCAGAGGTCATGAGgtca(-13bp)
F0-2 GGCCATGAGTGCTTTGGCATCGATGAGGCC(-4bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 ATGGGGAAAAAGATGATGCAGGAGGCCATGGCATCGATGAGGCCATGAGCTACGCAGAGG(-13bp)
F0-2 GGCCATGAGTGCTTTGGCATCGATGAGGCC(-4bp)
Allele name and predicted product 1020a
1020b
ATGGGGAAAAAGATGATGCAGGAGGCCATGGCATCGATGAGGCCATGAGCTACGCAGAGG(-13bp);truncated protein,135aa
GGCCATGAGTGCTTTGGCATCGATGAGGCC(-4bp);truncated protein,138aa
突变创建时间 2014-09-03
填表时间 2013/12/23
修改时间 2016/10/24
PI/负责人 (工作单位)
Genotype Information: