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ZKO number ZKO1091
Recent progress of this line F2 individuals have been indentified
Gene name sich211-202h22.9
Ensembl ID ENSDARG00000079980
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGATACGGTGCCGGCTGTACagg
Cas9 target site 2 sequence
Position in the gene Exon2
Restriction enzyme (RE) n/a
PCR primer (forward) CCGAAGATAACCCTGCCTAC
PCR primer (reverse) TTTAGAAGAACTCCTAGTGGGT
Length of expected PCR product (bp) 554
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 1091
Dosage of injected Cas9 mRNA (ng/ul) 400
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 10
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1091.pdf
Mutation efficiency in F0 embryos (%) 60
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 100% 4/4
Wild-type (WT) sequence TTAACTACCCTGTACAGCCGGCACCGTATCCACCTCCTGCAGGT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TTAACTACCCTGTAccCAGCCGGCACCGTATCCACCTCCTGCAGGT
F0-2 TTAACTACCCTGTA--GCCGGCACCGTATCCACCTCCTGCAGGT
Allele name and predicted product 1091a
1091b
TTAACTACCCTGTAccCAGCCGGCACCGTATCCACCTCCTGCAGGT Truncated protein (63aa)
TTAACTACCCTGTA--GCCGGCACCGTATCCACCTCCTGCAGGT Truncated protein (79aa)
突变创建时间 2013-11-05
填表时间 2013/12/20
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information: