If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO177
Recent progress of this line F1 individuals have been identified
Gene name cacng3a
Ensembl ID ENSDARG00000058460
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGACTCTGGAGACAGTGCTGCAtgg
Cas9 target site 2 sequence
Position in the gene 5', exon1
Restriction enzyme (RE) T7
PCR primer (forward) aactttactggcacgcgagagtt
PCR primer (reverse) tatgctggataagttggcggttc
Length of expected PCR product (bp) 553
Length of expected RE digestion product (bp) 250+303
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 177
Dosage of injected Cas9 mRNA (ng/ul) 200
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 70
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO177.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 20
Wild-type (WT) sequence tgtactcgcgcggagtctgcaggaccaagagcgtcaacgacaacgacacaaaccgcaagaatgaggaggtgctgacgcattcaggactctggagacagtgctgcatggaaggtactttttgtgttttcagaaagtagttaacatgcttttattgcatgtttaagggattcaacgttgtcttattaaactaaaaatgtcga
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 tgtactcgcgcggagtctgcaggaccaagagcgtcaacgacaacgacacaaaccgcaagaatgaggaggtgctgacgcattcaggactctggagacagtgcttgcatggaaggtactttttgtgttttcagaaagtagttaacatgcttttattgcatgtttaagggattcaacgttgtcttattaaactaaaaatgtcga (+1bp)
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 tgtactcgcgcggagtctgcaggaccaagagcgtcaacgacaacgacacaaaccgcaagaatgaggaggtgctgacgcattcaggactctggagacagtgcttgcatggaaggtactttttgtgttttcagaaagtagttaacatgcttttattgcatgtttaagggattcaacgttgtcttattaaactaaaaatgtcga (+1bp)
F0-2
Allele name and predicted product 177a
177b
truncated protein (77aa)
突变创建时间 2014-10-30
填表时间 2014/3/19
修改时间 2016/9/23
PI/负责人 (工作单位)
Genotype Information: