If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1058
Recent progress of this line F1 individuals have been identified
Gene name zgc:66455
Ensembl ID ENSDARG00000052375
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GATCCACATGCTTGCCCGG
Cas9 target site 2 sequence
Position in the gene Exon 3
Restriction enzyme (RE) Sma1/Xma1 (C︱CCGGG)
PCR primer (forward) gtgaatgttgcattaatgttgtg
PCR primer (reverse) gctctgtagcgtgcatagaag
Length of expected PCR product (bp) 571
Length of expected RE digestion product (bp) 129+403
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1058
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 91.6
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1058.pdf
Mutation efficiency in F0 embryos (%) 20
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 50% (2/4)
Wild-type (WT) sequence CCCCCGGGCAAGCATGTGGGATC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 CCCCCGGGGCAAGCATGTGGATC ( +1 bp) CCCCCG--CAAGCATGTGGATC ( -2 bp) CCCCCG---AAGCATGTGGATC (-3 bp)
F0-2 CCCCC--CTGCAACTGCAAGCATGTGGATC (+8 bp, -2 bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 25% (5/20) CCCCCGGGGCAAGCATGTGGATC (+1 bp)
F0-2 F1-2 10% (2/20) CCCCCG--CAAGCATGTGGATC ( -2 bp)
Allele name and predicted product 1058a
1058b
CCCCCGGGGCAAGCATGTGGATC; (+1) ;Truncated protein (111aa)
CCCCCG--CAAGCATGTGGATC ( -2 bp);Truncated protein (110aa)
突变创建时间 2013-09-10
填表时间 2014/1/4
修改时间 2016/11/11
PI/负责人 (工作单位)
Genotype Information: