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ZKO number ZKO1161
Recent progress of this line F2 generation has been raised
Gene name zgc:65894
Ensembl ID TUBB8P7 (1 of 3) ENSDARG00000016301
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGATCTAGAGCCTGGAACCA
Cas9 target site 2 sequence
Position in the gene exon3
Restriction enzyme (RE) n/a
PCR primer (forward) 5’- TCTGGGAGGTGATCAGCGATG -3’
PCR primer (reverse) 5’- CACCAAACTTGTTCCTGAAGATCC -3’
Length of expected PCR product (bp) 525
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids?used?for?making?gRNA pMD19T-gRNA
Primers?used?for?making?gRNA Pr-ZKO
Dosage of injected Cas9 mRNA (ng/ul) 600
Dosage of injected gRNA (ng/ul) 100
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency PCR and single clone sequencing
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1161.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 16.7%
Wild-type (WT) sequence TCCTTGTGGATCTAGAGCCTGGAACCATGGATTCA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 TCCT---------------------------------AA----ATGGCTCA(22bp deletion, 2bp insertion)
F0-2 TCCTTGTGGATCTAGAGCC-------ATGGATTCA(7bp deletion)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TCCT---------------------------------AA----ATGGCTCA(-20bp, 1bp mutation)
F0-2 TCCTTGTGGATCTAGAGCC-------ATGGATTCA(-7bp) TCTAGAGCCTGGATTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGACCT(30bp insertion, 7bp deletion)
Allele name and predicted product 1161a
1161b
GTGGAAAGTATGTCCCTCGTGCCATCCTAAATGGACTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGACCTGACAACTTTGTTTTTG
GTGGAAAGTATGTCCCTCGTGCCATCCTTGTGGATCTAGAGCCTGGATTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGACCTGACAACTTTGTTTTTG
突变创建时间
填表时间 2013/12/20
修改时间 2016/8/9
PI/负责人 (工作单位)
Genotype Information:
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