| ZKO number |
ZKO1166 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
mir722
|
|
Ensembl ID |
ENSDARG00000080893
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGACTGTGTTGCTGTCTGTGtgg
|
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Cas9 target site 2 sequence |
|
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Position in the gene |
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
gaggcctgtgactgtgtagag
|
|
PCR primer (reverse) |
ccagcaggactgacagagtta
|
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Length of expected PCR product (bp)
|
|
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Length of expected RE digestion product (bp) |
|
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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)
|
50
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
|
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Method to estimate mutation efficiency |
sequencing of PCR product
|
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Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1166.pdf
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Mutation efficiency in F0 embryos (%)
|
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Picture from F0 embryos
(gel picture or sequencing picture) |
|
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Mutation type
|
deletion
|
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Germline transmission efficiency in adult F0
|
22%
|
|
Wild-type (WT) sequence
|
gaggcctgtgactgtgtagaggtcaaagtatataggcatgagacattgtaactctaataaggatgtaagtgaagattaaacagcgctattttacatttgattattcaatttcggtgcctgaatgcaataaagtaaaatatttcacatatttcttcctaatactaatccagaataatccccattgtcttccagaataatcccaaatcgttccaaatagagttatctggtgaaactattcatatcgcaatcaaaacaaaatatagcagtgtcggtttgtcccagaatcctgcagccctaatcaggactgtgttgctgtctgtgtggctgcgcagatgttggggacgtccagcggagacctgtgtattggaacggAGTGGAATTTGAAACGTTTTGGCCAAAAATGTAGCCATGGCAAAGGGGTTTTTTGCAGAAACGTTTCAGATTTCGCTCCGTTCTccggcttcggaaaaaacctgctgctgccgagaggaagacgaagaggaacacatcatcactcatgtcgttgatactgtgtgtgtgtgtgtgtgagagagaggattctgtgtactgagggtcctggttttactttacttgaaaaattggtcaatgttcttgacatagactgaagtatgaagtgttgatgttttgctggacttcatgagttacagaaggggtttgattagtctttagtgattggctgatgtttctgtgagggcggagtttagggtcatagaaactctgatagaaaaacagtgcgaatctatgccatgtcctcactaagataggaaaacaaatgtgtgtgtgtgtgcatgtgacctgtgtgtatgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtttgtgtgtgtgtgtgtgtgatatgatggcttattaccagaggaatgtcagtgtgggaaatctgtggattgaatctcttccgctccgcagggattcagccagaaataggagtttactcttaacacacacacacacacacacacacacacacacacacacacacacacacacacacacttcgggtctgttcctgatgataatgatcatgctcatttctgagaggaaaacaatattgatcacttgatactgataactctgtcagtcctgctgg
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
gaggcctgtgactgtgtagaggtcaaagtatataggcatgagacattgtaactctaataaggatgtaagtgaagattaaacagcgctattttacatttgattattcaatttcggtgcctgaatgcaataaagtaaaatatttcacatatttcttcctaatactaatccagaataatccccattgtcttccagaataatcccaaatcgttccaaatagagttatctggtgaaactattcatatcgcaatcaaaacaaaatatagcagtgtcggtttgtcccagaatcctgcagccctaatcaggactgtgttgctgtcttccgcagggattcagccagaaataggagtttactcttaacacacacacacacacacacacacacacacacacacacacacacacacacacacacttcgggtctgttcctgatgataatgatcatgctcatttctgagaggaaaacaatattgatcacttgatactgataactctgtcagtcctgctgg
-667bp
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
gaggcctgtgactgtgtagaggtcaaagtatataggcatgagacattgtaactctaataaggatgtaagtgaagattaaacagcgctattttacatttgattattcaatttcggtgcctgaatgcaataaagtaaaatatttcacatatttcttcctaatactaatccagaataatccccattgtcttccagaataatcccaaatcgttccaaatagagttatctggtgaaactattcatatcgcaatcaaaacaaaatatagcagtgtcggtttgtcccagaatcctgcagccctaatcaggactgtgttgctgtcttccgcagggattcagccagaaataggagtttactcttaacacacacacacacacacacacacacacacacacacacacacacacacacacacacttcgggtctgttcctgatgataatgatcatgctcatttctgagaggaaaacaatattgatcacttgatactgataactctgtcagtcctgctgg
-667bp
|
|
F0-2
|
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Allele name and predicted product
|
1166a
1166b |
gaggcctgtgactgtgtagaggtcaaagtatataggcatgagacattgtaactctaataaggatgtaagtgaagattaaacagcgctattttacatttgattattcaatttcggtgcctgaatgcaataaagtaaaatatttcacatatttcttcctaatactaatccagaataatccccattgtcttccagaataatcccaaatcgttccaaatagagttatctggtgaaactattcatatcgcaatcaaaacaaaatatagcagtgtcggtttgtcccagaatcctgcagccctaatcaggactgtgttgctgtcttccgcagggattcagccagaaataggagtttactcttaacacacacacacacacacacacacacacacacacacacacacacacacacacacacttcgggtctgttcctgatgataatgatcatgctcatttctgagaggaaaacaatattgatcacttgatactgataactctgtcagtcctgctgg
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突变创建时间 |
2014-03-06
|
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填表时间 |
2013/12/17
|
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修改时间 |
2016/12/30
|
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PI/负责人 (工作单位) |
|