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ZKO number ZKO74
Recent progress of this line F1 progeny has been raised
Gene name kpnb3
Ensembl ID ENSDARG00000040245
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GATGGCCGTCTGGATGTCTAcgg
Cas9 target site 2 sequence
Position in the gene exon 3
Restriction enzyme (RE) Acc 1
PCR primer (forward) TTGTGGACTAACTATCAACCTG
PCR primer (reverse) AGTTTACGGTTGAGCATTGT
Length of expected PCR product (bp) 478
Length of expected RE digestion product (bp) 244+234
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 74
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 70
Method to estimate mutation efficiency Restriction enzyme digestionAcc 1
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO74.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
Wild-type (WT) sequence CTCATCTTTTGAGGAGGTTTATCCCAACCTGACCGTAGACATCCAGACGGCCATCAAGACAGAGCTGCTCGCTGGAATCCG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 CTCATCTTTTGAGGAGGTTTATCCCAAC-TGACCGTAGACATCCAGACGGCCATCAAGACAGAGCTGCTCGCTGGAATCCG (+0bp,-1bp)
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 20% CTCATCTTTTGAGGAGGTTTATCCCAAC-TGACCGTAGACATCCAGACGGCCATCAAGACAGAGCTGCTCGCTGGAATCCG (+0bp,-1bp)
F0-2
Allele name and predicted product 74a
74b
CTCATCTTTTGAGGAGGTTTATCCCAAC-TGACCGTAGACATCCAGACGGCCATCAAGACAGAGCTGCTCGCTGGAATCCG, Truncated protein (76aa)
突变创建时间 2015-06-18
填表时间 2013/12/18
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: