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ZKO number ZKO911
Recent progress of this line F2 individuals have been indentified
Gene name spryd7a
Ensembl ID ENSDARG00000052624
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAAATGCCCACGGTTCAGCtgg
Cas9 target site 2 sequence
Position in the gene exon1
Restriction enzyme (RE) PvuII(CAG︱CTG)
PCR primer (forward) GCCCCGTATGACAACTAAACT
PCR primer (reverse) TGATTTTGGTCCATTCCTGCT
Length of expected PCR product (bp) 521
Length of expected RE digestion product (bp) 163+358
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 (%) 75.3
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/ZKO911.pdf
Mutation efficiency in F0 embryos (%) 29.01
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type Indel
Germline transmission efficiency in adult F0 22.6
Wild-type (WT) sequence GGAAATGCCCACGGTTCAGCTGGACACTCATCACATGG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGAAATGCCCACGGTTCA--------CTCATCACATGG(-8bp)
F0-2 GGAAATGCCCACG-----GCTGGACACTCATCACATGG(-5bp)
Allele name and predicted product 911a
911b
MAALLSCCFGCCGDNNSGHVTLKEMPTVHSSHGTLITWVLMLSL-:;Truncated protein (44 aa)
MAALLSCCFGCCGDNNSGHVTLKEMPTAGHSSHGY-;Truncated protein (35 aa)
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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