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ZKO number ZKO1170
Recent progress of this line F2 individuals have been indentified
Gene name CABZ01059392.1
Ensembl ID ENSDARG00000063108
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GACGTAGTCAATCAATGTGGagg
Cas9 target site 2 sequence
Position in the gene exon2
Restriction enzyme (RE) n/a
PCR primer (forward) 5’- CTTTATCTCTGCTATCCTGGACCTC -3’
PCR primer (reverse) 5’- TTGTGCTCTTTTCCTTTACACTGTG -3’
Length of expected PCR product (bp) 481
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 1170
Dosage of injected Cas9 mRNA (ng/ul) 600
Dosage of injected gRNA (ng/ul) 100
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency PCR and single clone sequencing
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1170.pdf
Mutation efficiency in F0 embryos (%) 17
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 100*1/30%
Wild-type (WT) sequence TTACCGGAAGAGAGAAATTAACAAACAGAAAATGGAAGCTCTTTATGGAAAGTAGAAAGTTTCAGGACGTAGTCAATCAATGTGGAGGTAAATATCACGCTATCAACAGTAA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 TTACCGGAAGAGAGAAATTAACAAACAGAAAATGGAAGCTCTTTATGGAAAGTAGAAAGTTTCAGGACGTAGTCAATCAATGAGGTAAATATCACGCTATCAACAGTAA
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TTACCGGAAGAGAGAAATTAACAAACAGAAAATGGAAGCTCTTTATGGAAAGTAGAAAGTTTCAGGACGTAGTCAATCAATGAGGTAAATATCACGCTATCAACAGTAA
F0-2
Allele name and predicted product 1170a
1170b
TTACCGGAAGAGAGAAATTAACAAACAGAAAATGGAAGCTCTTTATGGAAAGTAGAAAGTTTCAGGACGTAGTCAATCAAT---GAGGTAAATATCACGCTATCAACAGTAA premature translation termination; Truncated protein (177aa)
突变创建时间 2015-03-02
填表时间 2013/12/20
修改时间 2016/8/9
PI/负责人 (工作单位)
Genotype Information: