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ZKO number ZKO846
Recent progress of this line F2 individuals have been indentified
Gene name p2rx3b
Ensembl ID ENSDARG00000029718
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGACTTCGTCAATCTCTGCA
Cas9 target site 2 sequence
Position in the gene Exon5
Restriction enzyme (RE) BanI
PCR primer (forward) GGCTTCAACTGTATATGCCACTG
PCR primer (reverse) GGTGAAGTTCTCCACCTCCA
Length of expected PCR product (bp) 434
Length of expected RE digestion product (bp) 294bp+140bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids?used?for?making?gRNA pMD19T-gRNA
Primers?used?for?making?gRNA Pr-ZKO 846
Dosage of injected Cas9 mRNA (ng/ul) 150
Dosage of injected gRNA (ng/ul) 175
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency Restriction enzyme digestionBanI
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO846.pdf
Mutation efficiency in F0 embryos (%) 60
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type Indel
Germline transmission efficiency in adult F0 33.3
Wild-type (WT) sequence GAGATCAAAGGATGGTGCCCTGCAGAGATTGACGAAGTCCAGCCGTAAGTCTGCATACAA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GAGATCAAAGGATGGTG - - - - - - - - AGATTGACGAAGTCCAGCCGTAAGTCTGCATACAA
F0-2 GAGATCAAAGGATGGTGCCCTG - - - - - ATTGACGAAGTCCAGCCGTAAGTCTGCATACAA
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GAGATCAAAGGATGGTG - - - - - - - - AGATTGACGAAGTCCAGCCGTAAGTCTGCATACAA
F0-2 GAGATCAAAGGATGGTGCCCTG - - - - - ATTGACGAAGTCCAGCCGTAAGTCTGCATACAA
Allele name and predicted product 846a
846b
(-8,+0)
(-5,+0)
突变创建时间 2014-03-19
填表时间 2013/12/19
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information:
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