If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO970
Recent progress of this line F2 individuals have been indentified
Gene name npnt
Ensembl ID ENSDARG00000018721
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGGCTGGACGAGAGTCTCCTggg
Cas9 target site 2 sequence
Position in the gene exon 2
Restriction enzyme (RE) n/a
PCR primer (forward) CTCTGTGCAGTGATTGGCTC
PCR primer (reverse) ATTGGTGACTTGCTGTGCTG
Length of expected PCR product (bp) 273
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 970
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 50
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO970.pdf
Mutation efficiency in F0 embryos (%) 50
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 100% (10/10)
Wild-type (WT) sequence GCATTGACTGCTGCTGGGGCTGGACGAGAGTCTCCTGGGGCCAGTGCCAGCGTGAGTACAGTAT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GCATTGACTGCTGCTGGGGCTGGACGAGAGTC----------AGTGCCAGCGTGAGTACAGTAT (-10bp)
F0-2 GCATTGACTGCTGCTGGGGCTGGACGAGAGTCTC(TGACA)CTGGGGCCAGTGCCAGCGTGAGTACAGTAT (+5bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 12.5%(1/8) GCATTGACTGCTGCTGGGGCTGGACGAGAGTC----------AGTGCCAGCGTGAGTACAGTAT (-10bp)
F0-2 27.3%(3/11) GCATTGACTGCTGCTGGGGCTGGACGAGAGTCTC(TGACA)CTGGGGCCAGTGCCAGCGTGAGTACAGTAT (+5bp)
Allele name and predicted product 970a
970b
GCATTGACTGCTGCTGGGGCTGGACGAGAGTC----------AGTGCCAGCGTGAGTACAGTAT Truncated protein(58aa)
GCATTGACTGCTGCTGGGGCTGGACGAGAGTCTC(TGACA)CTGGGGCCAGTGCCAGCGTGAGTACAGTAT Truncated protein(63aa)
突变创建时间 2014-10-16
填表时间 2013/12/19
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information: