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ZKO number ZKO644
Recent progress of this line F2 generation has been raised
Gene name klf5a
Ensembl ID ENSDARG00000015506
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGAGTCCTCCTGAAAGCCGTcgg
Cas9 target site 2 sequence
Position in the gene exon1
Restriction enzyme (RE) HinfI
PCR primer (forward) GTGTAGCCGCCTCACTTTCT
PCR primer (reverse) CGCACATTGCGAGACCATTT
Length of expected PCR product (bp) 566
Length of expected RE digestion product (bp) 319bp,237bp
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) 300
Dosage of injected gRNA (ng/ul) 35
Survival rate of F0 embryo @ 1dpf (%) 68
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO644.pdf
Mutation efficiency in F0 embryos (%) 15
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture)
Mutation type deletion
Germline transmission efficiency in adult F0 25
Wild-type (WT) sequence TTTCAGGAGGACTCCTCTCTGTTTTGCTTTGAACACAAGAATCCTGACTCGGAAAGGGGCTTATCCAACGA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 CTTAAACCCTCACTGTCCGACGGCTTTCAGGAGGACTCCTCTCTGTTTTGCTTT...........TCCTGACT -11bp
F0-2 CTTAAACCCTCA.......................................................CTGACT -55bp
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TTTCAGGAGGACTCCTCTCTGTTTTGCTTT-----------TCCTGACTCGGAAAGGGGCTTATCCAACGA (-11bp, +0bp)
F0-2 GATGACTTTCTCTCGCTTCTTAAACCCTCACTG-------------------------------------------------------ACTCGGAAAGGGGCTTATCCAACGACT (-55bp, +0bp)
Allele name and predicted product 644a
644b
TTTCAGGAGGACTCCTCTCTGTTTTGCTTT-----------TCCTGACTCGGAAAGGGGCTTATCCAACGA; truncated protein (41 aa)
GATGACTTTCTCTCGCTTCTTAAACCCTCACTG-------------------------------------------------------ACTCGGAAAGGGGCTTATCCAACGACT; truncated protein (41 aa)
突变创建时间 2013-08-28
填表时间 2013/12/23
修改时间 2016/10/24
PI/负责人 (工作单位)
Genotype Information:
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