| ZKO number |
ZKO1155 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
null
|
|
Ensembl ID |
CABZ01036821.1 ENSDARG00000086938
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
antisense strand
GGTCCTTACCAGTGGATAAC
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Intron1-Exon2
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
5’- TCCCTTAATTCTGATTGGCTG -3’
|
|
PCR primer (reverse) |
5’- TCCTGAAGACAAGCCTTTCCT -3’
|
|
Length of expected PCR product (bp)
|
400
|
|
Length of expected RE digestion product (bp) |
|
|
Plasmids used for making Cas9 mRNA |
pSP6-2sNLS-spCas9
|
| Plasmids used for making gRNA |
pT7-gRNA
|
| Primers used for making gRNA |
Pr-ZKO
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
670
|
|
Dosage of injected gRNA (ng/ul)
|
45
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
90
|
|
Method to estimate mutation efficiency |
PCR and single clone sequencing
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1155.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
91.7
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
50%(2/4: Number of positive F0 / Total number of F0 screened)
|
|
Wild-type (WT) sequence
|
tgaatgtagcGGTCCTTACCAGTGGATAACtggtggtcaaatc
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
tgaatgtagcGGTCCTTACCAGTGGAggtggtcaAACtggtggtcaaatc (+8 bp, -1 bp)
tgaatgtagcGGTCCTTACCAGTGGAxxxxxxxtggtcaaatc (-7 bp)
|
|
F0-2
|
tgaatgtagcGGTCCTTACCAGTGGAccttaccagaccttaccagtccttaccagtgAACtggtggtcaaatc (+31 bp, -1 bp)
tgaatgtagcGGTCCTTACCAGTGxxxxxxxxxxxgtcaaatc (-11 bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
78.95% (30/38): Number of F1 carrier / Total number of F1 screened)
tgaatgtagcGGTCCTTACCAGTGGATxxxxxxxggtcaaatc (-7 bp)
tgaatgtagcGGTCCTTACCAGTGGAggtggtcaAACtggtggtcaaatc (+8 bp, -1 bp)
tgaatgtagcGGTCCTTACCAGTGGATggcccttaccaAACtggtggtcaaatc (+11 bp)
tgaatgtagcGGTCCTTACCAGTGxxxAACtggtggtcaaatc (-3 bp)
tgaatgtagcGGTCCTTACCAGTGGATggtcaaaaccaAACtggtggtcaaatc (+11 bp)
tgaatgtagcGGTCCTTACCAGTGGxxxxxxxxxxxtcaaatc (-11 bp)
|
|
F0-2
|
87.5% (7/8): Number of F1 carrier / Total number of F1 screened)
tgaatgtagcGGTCCTTACCAGTGGxxAACtggtggtcaaatc
(-2 bp)
tgaatgtagcGGTCCTTACCAGTGGxxxxxxxxxxxtcaaatc (-11 bp)
tgaatgtagcGGTCCTTACCAxxxxxxxACtggtggtcaaatc
(-7 bp)
tgaatgtagcGGTCCTTACCAGTGGxxxxxxxxtggtcaaatc
(-8 bp)
tgaatgtagcGGTCCTTACCAGTGGAagtggtggtcaaatc
(-4 bp, +2 bp)
|
|
Allele name and predicted product
|
1155a
1155b |
tgaatgtagcGGTCCTTACCAGTGGATxxxxxxxggtcaaatc; Truncated protein (24 aa)
|
| tgaatgtagcGGTCCTTACCAGTGGAggtggtcaAACtggtggtcaaatc; Truncated protein (27 aa) |
|
突变创建时间 |
2014-06-04
|
|
填表时间 |
2013/12/20
|
|
修改时间 |
2016/8/9
|
|
PI/负责人 (工作单位) |
|