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ZKO number ZKO1002
Recent progress of this line F2 individuals have been indentified
Gene name etaa1
Ensembl ID ENSDARG00000074915
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGCAGTCAGGGTCATTGCTGggg
Cas9 target site 2 sequence
Position in the gene exon 2
Restriction enzyme (RE) Bsr DI
PCR primer (forward) ACTTGTTTGCATTTGTCAC
PCR primer (reverse) AGCCCTTAACTCAGATTTT
Length of expected PCR product (bp) 459
Length of expected RE digestion product (bp) 182+277
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1002
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/ZKO1002.pdf
Mutation efficiency in F0 embryos (%) 50
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0 12.5%(1/8)
Wild-type (WT) sequence CTGATCCCCGATGACTcccccagcaatgaccctgactgCCATCAG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 ATCCCCGATGACTCCCCCA----TGACCCTGACTGCCATCAGGACATTGTATG
F0-2 ATCCCCGATGACTCCCCCAG----------------------GACATTGTATGGGACACAACATCTCC
Allele name and predicted product 1002a
1002b
GACTCCCCCA----TGACCCTGACTGCCA trunacted protein(131aa)
ATCCCCGATGACTCCCCCAG----------------------GACATTGTATGGGACA trunacted protein(125aa)
突变创建时间 2013-12-15
填表时间 2013/12/19
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information: