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ZKO number ZKO489
Recent progress of this line F1 individuals have been identified
Gene name lrata
Ensembl ID ENSDARG00000037278
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
AATGCCCGACATCATGCCCC
Cas9 target site 2 sequence
Position in the gene Exon 1
Restriction enzyme (RE) NlaⅢ(CATG/)
PCR primer (forward) AGCCAGAGAAGGAGGAGTGT
PCR primer (reverse) ACCTGCAGTACGTCACGAAA
Length of expected PCR product (bp) 417
Length of expected RE digestion product (bp) 143+72+75+127
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids?used?for?making?gRNA pMD19T-gRNA
Primers?used?for?making?gRNA Pr-ZKO 489
Dosage of injected Cas9 mRNA (ng/ul) 320
Dosage of injected gRNA (ng/ul) 70
Survival rate of F0 embryo @ 1dpf (%) 85
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO489.pdf
Mutation efficiency in F0 embryos (%) 50
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type Indel
Germline transmission efficiency in adult F0 10% (1/10)
Wild-type (WT) sequence GCGAGAACAGAGTGGCTCACCTAATGCCCGACATCATGCCCCTGTTAACCAGCAACAAAC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GCGAGAACAGAGTGGCTCAC------------ATCATGCCCCTGTTAACCAGCAACAAAC
F0-2 GCGAGAACAGAGTGGCTCACCTAATGCC------------------------CAACAAAC
Allele name and predicted product 489a
489b
In-frame mutation. Lost L M P D
In-frame mutation. Lost D I M P L L T S
突变创建时间 2013-05-03
填表时间 2014/1/2
修改时间 2016/11/11
PI/负责人 (工作单位)
Genotype Information:
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