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ZKO number ZKO109
Recent progress of this line F2 individuals have been indentified
Gene name si:ch211-103i6.5
Ensembl ID ENSDARG00000074031
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GCGTGAGAAGATGCTGCTCagg
Cas9 target site 2 sequence
Position in the gene Exon 3
Restriction enzyme (RE) Dde I (CTN/AG)
PCR primer (forward) 5’- AACAAATCCGGTTTTACAGT -3’…………
PCR primer (reverse) 5’- TGCGAAACTCCCAGGTTGAT -3’…………
Length of expected PCR product (bp) 385
Length of expected RE digestion product (bp) 180 bp +205 bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 109
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 sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO109.pdf
Mutation efficiency in F0 embryos (%) 30
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0 30
Wild-type (WT) sequence AATGTTCTTGCCCCACGGATGTGTTGGATGCAGTAAAGCAGTTTCCAGCTTCACAGAATGACCTGAGCAGCATCTTCTCACGCATGCGGGAGAGCACCAAGAAGATGACTGCAGAGCAGCAGCG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 AATGTTCTTGCCCCACGGATGTGTTGGATGCAGTAAAGCAGTTTCCAGCTTCACAGAATGACCTG----------CATCTTCTCACGCATGCGGGAGAGCACCAAGAAGATGACTGCAGAGCAGCAGCG (+0bp, -5bp)
F0-2 AATGTTCTTGCCCCACGGATGTGTTGGATGCAGTAAAGCAGTTTCCAGCTTCACAGAATGACCT--------------------TCTCACGCATGCGGGAGAGCACCAAGAAGATGACTGCAGAGCAGCAGCG (+0bp, -11bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 AATGTTCTTGCCCCACGGATGTGTTGGATGCAGTAAAGCAGTTTCCAGCTTCACAGAATGACCTG----------CATCTTCTCACGCATGCGGGAGAGCACCAAGAAGATGACTGCAGAGCAGCAGCG (+0bp, -5bp)
F0-2 AATGTTCTTGCCCCACGGATGTGTTGGATGCAGTAAAGCAGTTTCCAGCTTCACAGAATGACCT--------------------TCTCACGCATGCGGGAGAGCACCAAGAAGATGACTGCAGAGCAGCAGCG (+0bp, -11bp)
Allele name and predicted product 109a
109b
ACCTG----------CATC truncated protein 137aa
ACCT--------------------TCTCA truncated protein 135aa
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: