If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1066
Recent progress of this line F1 individuals have been identified
Gene name cct4
Ensembl ID ENSDARG00000013475
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) Apa I (GGGCC|C)
Length of expected PCR product (bp) 471
Length of expected RE digestion product (bp) 72+399
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO
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 Restriction enzyme digestionRE digestion (BstYI)
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1066.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 20% (3/15)
Wild-type (WT) sequence tgccatcagaacaagtctgggccctaagggcatggataaaatggtatgttt
Germline mutations in selected F0
 (By screening F1 embryos)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 50% (m/n: Number of F1 carrier / Total number of F1 screened) tgccatcagaacaagtctgggcc---------tggataaaatggtatgtt (+0 bp, -9bp)
F0-2 27.3% (m/n: Number of F1 carrier / Total number of F1 screened) tgccatcagaacaa-------------gggcatggataaaatggtatgtt (+0 bp, -13 bp)
Allele name and predicted product 1066a
rdkpaqirysninagkavadairtslg---ldkmiqdgkgdvtitndgatilk; Truncated protein (530aa)
rdkpaqirysninagkavadairtrawik*; Truncated protein (79 aa)
突变创建时间 2014-02-15
填表时间 2014/1/4
修改时间 2016/11/11
PI/负责人 (工作单位)
Genotype Information: