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ZKO number ZKO181
Recent progress of this line F2 individuals have been indentified
Gene name ndufab1a
Ensembl ID ENSDARG00000058463
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GCAGCTCCGCTCATCCATGGCcgg
Cas9 target site 2 sequence
Position in the gene exon1
Restriction enzyme (RE) Hae III (GG/CC)
PCR primer (forward) tgctgctatgcactgagagt
PCR primer (reverse) agcgaaatgaggttgtttgt
Length of expected PCR product (bp) 416
Length of expected RE digestion product (bp) 272 +144
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 181
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 60
Method to estimate mutation efficiency Restriction enzyme digestion30
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO181.pdf
Mutation efficiency in F0 embryos (%) 30
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 30
Wild-type (WT) sequence CCATGGCCGGACACTTTCACATCTGACAACCGGACATAAGTCTTCACTGGCGCAG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 CCATGG TTTCACATCTGACAACC (+0bp,-8bp)
F0-2 CCATGG ACACTTTCACATCTGACAACCGG (+0bp,-4bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 CCATGG TTTCACATCTGACAACC (+0bp,-8bp)
F0-2
Allele name and predicted product 181a
181b
CCATGG------------ TTTCACATCTGACAACC truncated protein (44aa)
突变创建时间
填表时间 2013/12/17
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: