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ZKO number ZKO599
Recent progress of this line F2 individuals have been indentified
Gene name gli1
Ensembl ID ENSDARG00000033680
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAGTTCGTTTGCCACTGGAagg
Cas9 target site 2 sequence
Position in the gene Exon 9
Restriction enzyme (RE) n/a
PCR primer (forward) TTCGAGGCTTAAACCTGTTC
PCR primer (reverse) GTGCACTTGTGTGGTTTTTC
Length of expected PCR product (bp) 429
Length of expected RE digestion product (bp) n/a
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids?used?for?making?gRNA pT7-gRNA-ZKOpT7-gRNA-ZKO599
Primers?used?for?making?gRNA Pr-ZKO 599
Dosage of injected Cas9 mRNA (ng/ul) 200
Dosage of injected gRNA (ng/ul) 25
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO599.pdf
Mutation efficiency in F0 embryos (%) 33
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type Indel
Germline transmission efficiency in adult F0 8% (1/12)
Wild-type (WT) sequence AATGAGCACATCCACGGAGAGAAGAAGGAGTTCGTTTGCCACTGGAAGGACTGTTCCCGAGAGCAGCGTCCGTT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 AATGAGCACATCCACGGAGAGAAGAAGGAGAAGAAGGACTGTTCCCGAGAGCAGCGTCCGTT
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 AATGAGCACATCCACGGAGAGAAGAAGGAGAAGAAGGACTGTTCCCGAGAGCAGCGTCCGTT
F0-2
Allele name and predicted product 599a
599b
AATGAGCACATCCACGGAGAGAAGAAGGAGAAGAAGGACTGTTCCCGAGAGCAGCGTCCGTT delete aa335-338
突变创建时间 2014-07-11
填表时间 2013/12/18
修改时间 2016/9/21
PI/负责人 (工作单位)
Genotype Information:
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