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ZKO number ZKO1112
Recent progress of this line F2 individuals have been indentified
Gene name vps54
Ensembl ID ENSDARG00000002980
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGACCAGCTGCGCGAGACAGcgg
Cas9 target site 2 sequence
Position in the gene Exon 6
Restriction enzyme (RE) n/a
PCR primer (forward) TGGTGGAGGTGAGCATCG
PCR primer (reverse) TTTGGTGGTGGCGATGAG
Length of expected PCR product (bp) 326
Length of expected RE digestion product (bp) n/a
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids?used?for?making?gRNA pMD19T-gRNA
Primers?used?for?making?gRNA Pr-ZKO 1112
Dosage of injected Cas9 mRNA (ng/ul) 200
Dosage of injected gRNA (ng/ul) 100
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/ZKO1112.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 12.5% (2/16)
Wild-type (WT) sequence GCTGCAGGACCAGCTGCGCGAGACAGCGGACGCCGTGGCTAAAC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GCTGCAGGA-----------------------CCGTGGCTAAAC
F0-2 GCTGCAGGACCAGCTGCGCGAG--CGGACGCCCTGCGCGAGCGGACGCCGTGGCTAAAC
Allele name and predicted product 1112a
1112b
GCTGCAGGA-----------------------CCGTGGCTAAAC Truncated protein (254 aa)
GCTGCAGGACCAGCTGCGCGAG--CGGACGCCCTGCGCGAGCGGACGCCGTGGCTAAAC Truncated protein (266 aa)
突变创建时间 2014-01-21
填表时间 2013/12/20
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information:
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