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ZKO number ZKO907
Recent progress of this line F2 individuals have been indentified
Gene name rcbtb1
Ensembl ID ENSDARG00000036645
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAGCTCTCCTCTATTCGACagg
Cas9 target site 2 sequence
Position in the gene exon2
Restriction enzyme (RE) TaqI (T︱CGA)
PCR primer (forward) AAATACCAACCTCACCTCAGT
PCR primer (reverse) TCTCCAGTGCCCAGACAGTTAC
Length of expected PCR product (bp) 423
Length of expected RE digestion product (bp) 163+260
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids?used?for?making?gRNA pMD19T-gRNA
Primers?used?for?making?gRNA Pr-ZKO
Dosage of injected Cas9 mRNA (ng/ul) 500
Dosage of injected gRNA (ng/ul) 100
Survival rate of F0 embryo @ 1dpf (%) 14.3
Method to estimate mutation efficiency Restriction enzyme digestion利用软件ImageJ计算灰度
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO907.pdf
Mutation efficiency in F0 embryos (%) 31.87
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type deletion
Germline transmission efficiency in adult F0 30.8
Wild-type (WT) sequence GGAGCTCTCCTCTATTCGACAGGCAT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGAGCTCTCCTCT----GACAGGCAT(-4bp)
F0-2 GGAGCTCTCCTCTA-----CAGGCAT(-5bp)
Allele name and predicted product 907a
907b
MVDVSKWPLFTLLSAQELSSDRRHASLARLRMRPSTSPTMMKCTCWV-;Truncated protein(47aa)
MVDVSKWPLFTLLSAQELSSTGMHLWRVCE-;Truncated protein(30aa)
突变创建时间 2014-09-04
填表时间 2013/12/17
修改时间 2016/8/12
PI/负责人 (工作单位)
Genotype Information:
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