If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO977
Recent progress of this line F2 generation has been raised
Gene name pkd2
Ensembl ID ENSDARG00000014098
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGACAACCCAGGGTTCGAGCcgg
Cas9 target site 2 sequence
Position in the gene exon 1
Restriction enzyme (RE) n/a
PCR primer (forward) ATCTGCATCCGCATCTCC
PCR primer (reverse) CAGTATCCTCTTCCCACA
Length of expected PCR product (bp) 343
Length of expected RE digestion product (bp) 144+199
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 977
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 10
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO977.pdf
Mutation efficiency in F0 embryos (%) 60
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0 67.5%(5/8)
Wild-type (WT) sequence GACAACCCAGGGTTCGAGCCGGAGGAGGGGATGATGGA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GACAACCCAGGGTT-----CGGAGGAGGGGATGAT
F0-2 GACAACCCAGGGT----GGAGGAGGAGGGATG
Allele name and predicted product 977a
977b
GACAACCCAGGGTT-----CGGAGGAGGGGATGA truncated protein(142aa)
GGGACAACCCAGGGT----GGAGGAGGAGGGATG truncated protein(71aa)
突变创建时间
填表时间 2013/12/19
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information: