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ZKO number ZKO204
Recent progress of this line F2 individuals have been indentified
Gene name pdia2
Ensembl ID ENSDARG00000018263
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GAACAAATTTTCAACTCCAagg
Cas9 target site 2 sequence
Position in the gene exon6
Restriction enzyme (RE) Sty I
PCR primer (forward) GGTCGTTCCATTCAACGAGGA
PCR primer (reverse) CTGGCAAAACGTTCTCAGTG
Length of expected PCR product (bp) 548
Length of expected RE digestion product (bp) 181+367
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 204
Dosage of injected Cas9 mRNA (ng/ul) 400
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency Restriction enzyme digestionSty I
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO204.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 40%
Wild-type (WT) sequence aattaacatttattttgcagAATGCCGAACAAATTTTCAACTCCAAGGTCCGTAAGCATTTGCTTCTATTTCTTAATACAACAGTGGATTCCCAGAATGCATTAGTAGAGGAGTTTAGAGAGGTTGCCAGTGAGTTTAAGGAGAAG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 aattaacatttattttgcagAATGCCGAACAAATTTTCAACCAAGGTCCGTAAGCATTTGCTTCTATTTCTTAATACAACAGTGGATTCCCAGAATGCATTAGTAGAGGAGTTTAGAGAGGTTGCCAGTGAGTTTAAGGAGAAG -2bp
F0-2 aattaacatttattttgcagAATGCCGAACAAATTTTAAGGTCCGTAAGCATTTGCTTCTATTTCTTAATACAACAGTGGATTCCCAGAATGCATTAGTAGAGGAGTTTAGAGAGGTTGCCAGTGAGTTTAAGGAGAAG -7bp
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 aattaacatttattttgcagAATGCCGAACAAATTTTCAACCAAGGTCCGTAAGCATTTGCTTCTATTTCTTAATACAACAGTGGATTCCCAGAATGCATTAGTAGAGGAGTTTAGAGAGGTTGCCAGTGAGTTTAAGGAGAAG -2bp
F0-2 aattaacatttattttgcagAATGCCGAACAAATTTTAAGGTCCGTAAGCATTTGCTTCTATTTCTTAATACAACAGTGGATTCCCAGAATGCATTAGTAGAGGAGTTTAGAGAGGTTGCCAGTGAGTTTAAGGAGAAG -7bp
Allele name and predicted product 204a
204b
aattaacatttattttgcagAATGCCGAACAAATTTTCAACCAAGGTCCGTAAGCATTTGCTTCTATTTCTTAATACAACAGTGGATTCCCAGAATGCATTAGTAGAGGAGTTTAGAGAGGTTGCCAGTGAGTTTAAGGAGAAG
aattaacatttattttgcagAATGCCGAACAAATTTTAAGGTCCGTAAGCATTTGCTTCTATTTCTTAATACAACAGTGGATTCCCAGAATGCATTAGTAGAGGAGTTTAGAGAGGTTGCCAGTGAGTTTAAGGAGAAG
突变创建时间 2014-06-23
填表时间 2013/12/17
修改时间 2016/12/30
PI/负责人 (工作单位)
Genotype Information: