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ZKO number ZKO175
Recent progress of this line F2 generation has been raised
Gene name tlr2
Ensembl ID ENSDARG00000037758
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGGTGGGAGACATCTTGAagg
Cas9 target site 2 sequence
Position in the gene exon2
Restriction enzyme (RE) T7
PCR primer (forward) ggtcatatgagaacgggagcttg
PCR primer (reverse) aagtaggctgtgcgtaggctgtc
Length of expected PCR product (bp) 385
Length of expected RE digestion product (bp) 105+280
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 175
Dosage of injected Cas9 mRNA (ng/ul) 200
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO175.pdf
Mutation efficiency in F0 embryos (%) 20
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0 30
Wild-type (WT) sequence ggtcatatgagaacgggagcttgaaagcagctcgtcccatcggttcagtctctttgagtctacagaatttgtttgagagtgatccagaactagtttccaaggtccttcaagatgtctcccaccctgaaactctgctgataatcaaagatgtcactatgaaaacaaacacatctacagaacccttcaaaatggtgaaggaaggtggcactaagagcctgactttccaaaactcctccacgaccgatcaagccttgacctcttttttagagttcatggatggctctccattgtcttttattggacttgaagatatacaatttgttgggataggagagtggcagaaagctaaatacacacatcatgacagcctacgcacagcctactt
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 ggtcatatgagaacgggagcttgaaagcagctcgtcccatcggttcagtctctttgagtctacagaatttgtttgagagtgatccagaactagtttccaaggtcctagtttccaagatgtctcccaccctgaaactctgctgataatcaaagatgtcactatgaaaacaaacacatctacagaacccttcaaaatggtgaaggaaggtggcactaagagcc (+5bp)
F0-2 GTCTCTTTGAGTCTACAGAATTTGTTTGAGAGTGATCCAGAACTAGTTTCCAAGGTCCAAACAAACACATCTACAGAACCCTTCAAAATGGTGAAGGAAGGTGGCACTAAGAGCCTGACTTTCCAAAACTCCT (-54bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 ggtcatatgagaacgggagcttgaaagcagctcgtcccatcggttcagtctctttgagtctacagaatttgtttgagagtgatccagaactagtttccaaggtcctagtttccaagatgtctcccaccctgaaactctgctgataatcaaagatgtcactatgaaaacaaacacatctacagaacccttcaaaatggtgaaggaaggtggcactaagagcc (+5bp)
F0-2 GTCTCTTTGAGTCTACAGAATTTGTTTGAGAGTGATCCAGAACTAGTTTCCAAGGTCCAAACAAACACATCTACAGAACCCTTCAAAATGGTGAAGGAAGGTGGCACTAAGAGCCTGACTTTCCAAAACTCCT (-54bp)
Allele name and predicted product 175a
175b
truncated protein (260aa)
deletion (-18aa)
突变创建时间
填表时间 2014/3/19
修改时间 2016/9/23
PI/负责人 (工作单位)
Genotype Information: