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ZKO number ZKO105
Recent progress of this line F2 individuals have been indentified
Gene name cnppd1
Ensembl ID ENSDARG00000027078
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GCTCTATCATGCCCCATGACagg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) Bsl I (CCNN/NNNNNGG)
PCR primer (forward) 5’- tgttactgaacgaatgtgcga -3’…………51.7
PCR primer (reverse) 5’- ggacaacagtatctcccgcat -3’…………53.0
Length of expected PCR product (bp) 608
Length of expected RE digestion product (bp) 266 bp + 342 bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 105
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 75
Method to estimate mutation efficiency Restriction enzyme digestionBsl 1
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO105.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 30% 3/10
Wild-type (WT) sequence ACAGCACGCTGGATGCTCTATCATGCCCCATGACAGgtgagtcaacttaatttaaaaataggcatgg
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 ACGCTGGATGCTCTATCATGCCGTGACAGgtgagtcaacttaatttaaaaataggcatgg (+1bp, -3 bp)
F0-2 ACAGCACGCTGGATGCTCTATCATGCCTAAGTCGACAGgtgagtcaacttaatttaaaaataggcatgg (-4bp, +6bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 ACGCTGGATGCTCTATCATGCCGTGACAGgtgagtcaacttaatttaaaaataggcatgg (+1bp, -3 bp)
F0-2 ACAGCACGCTGGATGCTCTATCATGCCTAAGTCGACAGgtgagtcaacttaatttaaaaataggcatgg (-4bp, +6bp)
Allele name and predicted product 105a
105b
ATCATGCC--GTGACAG truncated protein(69aa)
ATCATGCCTAAGTCGACAG(59aa)
突变创建时间
填表时间 2013/12/16
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: