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ZKO number ZKO1155
Recent progress of this line F2 generation has been raised
Gene name null
Ensembl ID CABZ01036821.1 ENSDARG00000086938
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
Cas9 target site 2 sequence
Position in the gene Intron1-Exon2
Restriction enzyme (RE) n/a
PCR primer (forward) 5’- TCCCTTAATTCTGATTGGCTG -3’
PCR primer (reverse) 5’- TCCTGAAGACAAGCCTTTCCT -3’
Length of expected PCR product (bp) 400
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pSP6-2sNLS-spCas9
Plasmids used for making gRNA pT7-gRNA
Primers used for making gRNA Pr-ZKO
Dosage of injected Cas9 mRNA (ng/ul) 670
Dosage of injected gRNA (ng/ul) 45
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/ZKO1155.pdf
Mutation efficiency in F0 embryos (%) 91.7
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 50%(2/4: Number of positive F0 / Total number of F0 screened)
Wild-type (WT) sequence tgaatgtagcGGTCCTTACCAGTGGATAACtggtggtcaaatc
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 tgaatgtagcGGTCCTTACCAGTGGAggtggtcaAACtggtggtcaaatc (+8 bp, -1 bp) tgaatgtagcGGTCCTTACCAGTGGAxxxxxxxtggtcaaatc (-7 bp)
F0-2 tgaatgtagcGGTCCTTACCAGTGGAccttaccagaccttaccagtccttaccagtgAACtggtggtcaaatc (+31 bp, -1 bp) tgaatgtagcGGTCCTTACCAGTGxxxxxxxxxxxgtcaaatc (-11 bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 78.95% (30/38): Number of F1 carrier / Total number of F1 screened) tgaatgtagcGGTCCTTACCAGTGGATxxxxxxxggtcaaatc (-7 bp) tgaatgtagcGGTCCTTACCAGTGGAggtggtcaAACtggtggtcaaatc (+8 bp, -1 bp) tgaatgtagcGGTCCTTACCAGTGGATggcccttaccaAACtggtggtcaaatc (+11 bp) tgaatgtagcGGTCCTTACCAGTGxxxAACtggtggtcaaatc (-3 bp) tgaatgtagcGGTCCTTACCAGTGGATggtcaaaaccaAACtggtggtcaaatc (+11 bp) tgaatgtagcGGTCCTTACCAGTGGxxxxxxxxxxxtcaaatc (-11 bp)
F0-2 87.5% (7/8): Number of F1 carrier / Total number of F1 screened) tgaatgtagcGGTCCTTACCAGTGGxxAACtggtggtcaaatc (-2 bp) tgaatgtagcGGTCCTTACCAGTGGxxxxxxxxxxxtcaaatc (-11 bp) tgaatgtagcGGTCCTTACCAxxxxxxxACtggtggtcaaatc (-7 bp) tgaatgtagcGGTCCTTACCAGTGGxxxxxxxxtggtcaaatc (-8 bp) tgaatgtagcGGTCCTTACCAGTGGAagtggtggtcaaatc (-4 bp, +2 bp)
Allele name and predicted product 1155a
tgaatgtagcGGTCCTTACCAGTGGATxxxxxxxggtcaaatc; Truncated protein (24 aa)
tgaatgtagcGGTCCTTACCAGTGGAggtggtcaAACtggtggtcaaatc; Truncated protein (27 aa)
突变创建时间 2014-06-04
填表时间 2013/12/20
修改时间 2016/8/9
PI/负责人 (工作单位)
Genotype Information: