If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO107
Recent progress of this line F2 individuals have been indentified
Gene name fam134a
Ensembl ID ENSDARG00000063270
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGGTCTGGAGGCCTGGTTCCcgg
Cas9 target site 2 sequence
Position in the gene Exon1
Restriction enzyme (RE) Stu I(AGG/CCT)
PCR primer (forward) 5’- tccatgacgagcctctcct -3’…………51.8
PCR primer (reverse) 5’- accacggtaagagtagcgct -3’…………50.9
Length of expected PCR product (bp) 438
Length of expected RE digestion product (bp) 120bp + 318bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 107
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 63
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO107.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 2/15
Wild-type (WT) sequence GAGGCCTGGTTCCCGGCGCCCGGTGGCGCGGAGGAGGAACCGGAG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GAGGCCTGGTTCCCGGCGCCCGG GAGGCC------------------------GCCCGG (+0bp,-11bp)
F0-2 GAGGCCTGGTTCCCGGCGCCCGGTGGCGCGGAGGAGGAACCGGAGCT GAGG----------------------------------------------------------------------------AACCGGAGCT (+0bp, -33bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GAGGCC------------------------GCCCGG (+0bp,-11bp)
F0-2 GAGG----------------------------------------------------------------------------AACCGGAGCT (+0bp, -33bp)
Allele name and predicted product 107a
107b
GAGGCC------------------------GCCCGG Truncated protein (115aa)
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: