| ZKO number |
ZKO1075 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
acta1a
|
|
Ensembl ID |
ENSDARG00000036371
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
antisense strand
GGCTCTAGGGGCGTCATCTCcgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon2
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
ACAGCTGAATGAATGCTTAAGTCT
|
|
PCR primer (reverse) |
ACCCATCTTCAAGACACTTCT
|
|
Length of expected PCR product (bp)
|
594
|
|
Length of expected RE digestion product (bp) |
n/a
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids used for making gRNA |
pMD19T-gRNA
|
| Primers used for making gRNA |
Pr-ZKO 1075
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
100
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
75
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1075.pdf
|
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Mutation efficiency in F0 embryos (%)
|
50
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
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Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
67%
|
|
Wild-type (WT) sequence
|
ACAGCTGAATGAATGCTTAAGTCTATTTACCTGATTATATTTGAATTACATCACAACATCGATGCTGTTAAAGAAACCATATGAAATTGAAAATAGTCACATAAACCGTTTACCGGAAGTTTATCAGTGGCTGAAAATCATTAGCTGTGCATATATAGACCATTTTTAATGAAGTTTGTGTGGATTTGTTTTCTCACCTGCCTTGTCTGTTTGTAGAACCCATCAAGATGTGTGACGATGATGAGACCACAGCTTTGGTGTGCGACAACGGATCAGGCCTGGTCAAGGCTGGTTTCGCCGGAGATGACGCCCCTAGAGCCGTCTTCCCATCCATTGTCGGCCGCCCGCGTCACCAGGTAAAATCCTGAAATATGAAATAAAGGTCTCGTGAAGTGCTTTGAAATGTGCAATTTTAATTCGATGTTTGACATAGACTTAACTGAAAAATGAAGAGAGTGCAGGGCAAAGAGTTGCTCCTCCCCTTTTTAAAAAGCAGCCAATAGCATTTTGTTTTTCTCACAGCTCTGCCAGTGAGAGTAGTTGAGCTCAAGCGCATTAAATGAAAAGCAAATAAGAAGTGTCTTGAAGATGGGT
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
-24
ACAGCTGAATGAATGCTTAAGTCTATTTACCTGATTATATTTGAATTACATCACAACATCGATGCTGTTAAAGAAACCATATGAAATTGAAAATAGTCACATAAACCGTTTACCGGAAGTTTATCAGTGGCTGAAAATCATTAGCTGTGCATATATAGACCATTTTTAATGAAGTTTGTGTGGATTTGTTTTCTCACCTGCCTTGTCTGTTTGTAGAACCCATCAAGATGTGTGACGATGATGAGACCACAGCTTTGGTGTGCGACAACGGATCAGGCCTGGTCAAGGCTGGTTTCG---------------------TCTTCCCATCCATTGTCGGCCGCCCGCGTCACCAGGTAAAATCCTGAAATATGAAATAAAGGTCTCGTGAAGTGCTTTGAAATGTGCAATTTTAATTCGATGTTTGACATAGACTTAACTGAAAAATGAAGAGAGTGCAGGGCAAAGAGTTGCTCCTCCCCTTTTTAAAAAGCAGCCAATAGCATTTTGTTTTTCTCACAGCTCTGCCAGTGAGAGTAGTTGAGCTCAAGCGCATTAAATGAAAAGCAAATAAGAAGTGTCTTGAAGATGGGT
|
|
F0-2
|
-3, +20
ACAGCTGAATGAATGCTTAAGTCTATTTACCTGATTATATTTGAATTACATCACAACATCGATGCTGTTAAAGAAACCATATGAAATTGAAAATAGTCACATAAACCGTTTACCGGAAGTTTATCAGTGGCTGAAAATCATTAGCTGTGCATATATAGACCATTTTTAATGAAGTTTGTGTGGATTTGTTTTCTCACCTGCCTTGTCTGTTTGTAGAACCCATCAAGATGTGTGACGATGATGAGACCACAGCTTTGGTGTGCGACAACGGATCAGGCCTGGTCAAGGCTGGTTTCGCCGGA aaactcggtttcaaaactcgGACGCCCCTAGAGCCGTCTTCCCATCCATTGTCGGCCGCCCGCGTCACCAGGTAAAATCCTGAAATATGAAATAAAGGTCTCGTGAAGTGCTTTGAAATGTGCAATTTTAATTCGATGTTTGACATAGACTTAACTGAAAAATGAAGAGAGTGCAGGGCAAAGAGTTGCTCCTCCCCTTTTTAAAAAGCAGCCAATAGCATTTTGTTTTTCTCACAGCTCTGCCAGTGAGAGTAGTTGAGCTCAAGCGCATTAAATGAAAAGCAAATAAGAAGTGTCTTGAAGATGGGT
|
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Allele name and predicted product
|
1075a
1075b |
AGGCTGGTTTCG-------------------------------------------TCTTCCCATCCAT（Truncated protein 368aa）
|
| GGTTTCGCCGGA---[aaactcggtttcaaaactcg]中括号内为插入碱基GACGCCCCTA（Truncated protein 25aa） |
|
突变创建时间 |
2015-04-18
|
|
填表时间 |
2013/12/17
|
|
修改时间 |
2016/9/23
|
|
PI/负责人 (工作单位) |
|