If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO969
Recent progress of this line F2 individuals have been indentified
Gene name gstcd
Ensembl ID ENSDARG00000036494
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GCCAGTTGATGCCCGACCGCAggg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) Bsl I (CCNN/NNNNNGG)
PCR primer (forward) TGTCCTACAGCGAATGCTC
PCR primer (reverse) ctaagcatttccagcacctgt
Length of expected PCR product (bp) 369
Length of expected RE digestion product (bp) 250 bp +119 bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 969
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 52
Method to estimate mutation efficiency Restriction enzyme digestion68
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO969.pdf
Mutation efficiency in F0 embryos (%) 30
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0 6/20
Wild-type (WT) sequence CCCGACCGCAGGGACGTGGCGTCACTTTT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 CCCGACC AGGGACGTGGCGTCACTTTT(+0bp,-2bp)
F0-2 CCCG-------CAGGGACGTGGCG (+0bp,-4bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 CCCGACC AGGGACGTGGCGTCACTTTT(+0bp,-2bp)
F0-2 CCCG-------CAGGGACGTGGCG (+0bp,-4bp)
Allele name and predicted product 969a
969b
GCCAGTTGATGCCCGACC-- AGGGAC truncated protein (128aa)
CCCG---CAGGGACGTGGCG truncated protein (153aa)
突变创建时间 2015-03-18
填表时间 2013/12/17
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: