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ZKO number ZKO1157
Recent progress of this line F2 individuals have been indentified
Gene name si:dkey-76b14.5
Ensembl ID C3 (5 of 9) ENSDARG00000053462
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGGATGTCCTTCGATGTCTC
Cas9 target site 2 sequence
Position in the gene exon9
Restriction enzyme (RE) n/a
PCR primer (forward) 5’- AGGCAGTGAAATGGTGGATG -3’
PCR primer (reverse) 5’- TTTAGTGATGCCGTTGGGTT -3’
Length of expected PCR product (bp) 302
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
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/ZKO1157.pdf
Mutation efficiency in F0 embryos (%) 80
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type Indel
Germline transmission efficiency in adult F0 50%
Wild-type (WT) sequence AACCCGGGATGTCCTTCGATGTCTCGGTAAAA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 AA---------TCCTTCGATGTCTC (-9 bp)
F0-2 AAACCCGGGTCCTTCAAACTTCAGTCCTTCGATGTCTCGGTAAAA (+14bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 CAAACCCGGCAATTCTTCATCCTTCGATGTCTCGGTAAAA(+ 6bp,3bp mutation)
F0-2 CAAACCCGGAGAATTCGTCCTTCGATGTCTCGGTAAA (+4bp, 1bp mut)
Allele name and predicted product 1157a
1157b
CAAACCCGGAGAATTCGTCCTTCGATGTCTCGGTAAA (+4bp,1bp mutant)truncated protein
CAAACCCGGCAATTCTTCATCCTTCGATGTCTCGGTAAAA(+ 6bp,3bp mutation)truncated protein
突变创建时间 2014-04-18
填表时间 2013/12/20
修改时间 2016/8/9
PI/负责人 (工作单位)
Genotype Information:
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