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ZKO number ZKO1059
Recent progress of this line F2 generation has been raised
Gene name b3gnt2a
Ensembl ID ENSDARG00000052376
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGACGAGCTGGAATCCTCGAtgg
Cas9 target site 2 sequence
Position in the gene exon1
Restriction enzyme (RE) n/a
PCR primer (forward) GCCTCCTCATTTTCGGGGAA
PCR primer (reverse) TCGCCGTATAACTCCGCTTC
Length of expected PCR product (bp) 378
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 45
Survival rate of F0 embryo @ 1dpf (%) 86
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1059.pdf
Mutation efficiency in F0 embryos (%) 15
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0
Wild-type (WT) sequence TTCGGGAATCGTGGGGACGAGCTGGAATCCTCGATGGTCAGCGCATTGCTACAGTTTTCTT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 TTCGGGAATCGTGGGGACGAGCTGGAATCCGTCCAGCGCATTGCTACAGTTTTCTT -7bp
F0-2 TTCGGGAATCGTGGGGACGAGCTGGAATCCTCGGTCAGCGCATTGCTACAGTTTTCTT
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TTCGGGAATCGTGGGGACGAGCTGGAATCCGTCCAGCGCATTGCTACAGTTTTCTT(-8bp,+3bp
F0-2 TTCGGGAATCGTGGGGACGAGCTGGAATCCTC…GGTCAGCGCATTGCTACAGTTTTCTT( -3 bp)
Allele name and predicted product 1059a
1059b
TTCGGGAATCGTGGGGACGAGCTGGAATCCGTCCAGCGCATTGCTACAGTTTTCTT(219AA)turncate protein
TTCGGGAATCGTGGGGACGAGCTGGAATCCTC…GGTCAGCGCATTGCTACAGTTTTCTT(-1aa)
突变创建时间 2013-07-08
填表时间 2013/12/23
修改时间 2016/10/24
PI/负责人 (工作单位)
Genotype Information: