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ZKO number ZKO1172
Recent progress of this line F2 generation has been raised
Gene name zgc:171452
Ensembl ID ENSDARG00000074069
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAGGAGCCATTGAAGGAGCagg
Cas9 target site 2 sequence
Position in the gene exon2
Restriction enzyme (RE) n/a
PCR primer (forward) 5’- ATGCTGGAGGAAAACGGAGG -3’
PCR primer (reverse) 5’- GCATCACTCCAACACTGTCC -3’
Length of expected PCR product (bp) 323
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1172
Dosage of injected Cas9 mRNA (ng/ul) 667
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1172.pdf
Mutation efficiency in F0 embryos (%) 30
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0 44%
Wild-type (WT) sequence GGAGGAGCCATTGAAGGAGCagg
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GGAGGAGCCATTGAAGGAGCxxx (+0 bp, -3 bp)
F0-2 GGxxxxxxxxxxxxAGGAGCagg (+0 bp, -12 bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 10%( 2/20: Number of positive F1 / Total number of F1 screened ) GGAGGAACCACAGGAAACGCTCCTTCAATGGCTCCTCCTTTGAAGGA (+41bp, -24bp)
F0-2 GGAGGAGCCATTGAxxxAGCagg (+0bp, -3bp)
Allele name and predicted product 1172a
1172b
ATGTCAGACCTCAGGATTGTAATGTTGGGTGAAAGTGGAGCAGGGAAGAGCTCATCAGGA AACACCATCCTGGGAGAAAAAGTGTTCATTAAACAATTCACTGATAAATCTGTGACCGAG AAATGCCAGAAACACCAGCGTGAGGTCAAAGGTCGGATCATCTCCGTCATCGACACTCCA GGACTGTGTGACACGTCGATAAATAAAGAAGAAGTGAAGAAAGAAATGGAGAAAAGCACT GAAACATCTGCTCCTGGTCCTCACGTGTTTCTGCTGGTGCTCAGACTGGACGAGAAACCA GCAAACCAAGAGAAAAACACAATGAAATGGATCCAGGAGAACTTTGGGGAAGAAGCTAAT CGGTACACCATCATCCTCTTCACTAGAGGAGATCAGATTAAAACATCCATAGAGGAGTTT CTGGCAAATAACGAGGAGATGAGAGCACTAGCTGAACAGTGTAAAGGTGGATACCATGTC TTCAACAACACAGATGAACAAAACAGATCTCAGGTCAGTGAGCTGCTGGAGAAGATCGAC TCAATGCTGGAGGAAAACGGAGGACAGTTTTACACCAATGAGATGTACATGGAGGCTCAG AGAGGCAGAAATCTGACCAGGGCTCAGAAAGCGGTGTTTGTGGGAGCAGGT CTGCTTGGAGGAACCACAGGAAACGCTCCTTCAATGGCTCCTCCTTTGAAGGAGCTGTGGTTCAAACTGCCATGGCTTTAG Truncated protein (244aa)
突变创建时间 2014-09-11
填表时间 2013/12/20
修改时间 2016/8/9
PI/负责人 (工作单位)
Genotype Information: