| ZKO number |
ZKO22 |
| Recent progress of this line |
F0 population has been raised
|
|
Gene name |
trmt10c
|
|
Ensembl ID |
ENSDARG00000041575
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGTGTTTCCCGCCGGTCAGAcgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 2
|
|
Restriction enzyme (RE) |
T7E1
|
|
PCR primer (forward) |
AGTCTCTTAAATCCCCTTTCTTCCCCATTC
|
|
PCR primer (reverse) |
TAAGTGTAGCATCACCTTTCCAGCTTG
|
|
Length of expected PCR product (bp)
|
533
|
|
Length of expected RE digestion product (bp) |
293+240
|
|
Plasmids used for making Cas9 mRNA |
pSP6-2sNLS-spCas9
|
| Plasmids used for making gRNA |
pT7-gRNA-ZKOpT7-gRNA-ZKO21
|
| Primers used for making gRNA |
Pr-ZKO 22
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
35
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
50
|
|
Method to estimate mutation efficiency |
T7EI detection
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO22.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
100
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
50
|
|
Wild-type (WT) sequence
|
CTGATGATGTTTCTGCGCTGGTGTTTCCCGCCGGTCAGACGGATGTGTGTGTGTCAGCAGATCTCCGTCAGGTCCTTCAGCAGCAGCCGGACGCTCCTCAAAGATGACC
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
CTGATGATGTTTCTGCGCTGGTGTTTCCCGCCGG--------ATGTGTGTGTGTCAGCAGATCTCCGTCAGGTCCTTCAGCAGCAGCCGGACGCTCCTCAAAGATGACC(-8bp,+0bp)
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
CTGATGATGTTTCTGCGCTGGTGTTTCCCGCCGG--------ATGTGTGTGTGTCAGCAGATCTCCGTCAGGTCCTTCAGCAGCAGCCGGACGCTCCTCAAAGATGACC(-8bp,+0bp)
|
|
F0-2
|
|
|
Allele name and predicted product
|
22a
22b |
TGCGCTGGTGTTTCCCGCCGG--------ATGTGTGTGTGTCAGCAGATCTCCGTCAGGTCCTTCAGCAGCA Truncated Protein (31aa)
|
| |
|
突变创建时间 |
2014-11-25
|
|
填表时间 |
2013/12/24
|
|
修改时间 |
2016/8/8
|
|
PI/负责人 (工作单位) |
|