| ZKO number |
ZKO998 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
atg4d
|
|
Ensembl ID |
ENSDARG00000043548
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGCCATGGGGCGGTTCCCGAcgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon2
|
|
Restriction enzyme (RE) |
NcoI
|
|
PCR primer (forward) |
ACCTGTCCTCAGTCAGGTGT
|
|
PCR primer (reverse) |
GTTGTTCCAGGCCGACATGA
|
|
Length of expected PCR product (bp)
|
651
|
|
Length of expected RE digestion product (bp) |
445bp,206bp
|
|
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 (%)
|
85
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestionNcoI
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO998.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
25
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
34
|
|
Wild-type (WT) sequence
|
GTTCAGTATATAGTGGGTGGTGGAGCTCATGAGGACAAGGCTTCAGCTTCGTCCAAACGC
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
AACGCCACCTTGGCCATGGGGCGGTTCCCGAC..........AAGGTTCTGGA -10bp
|
|
F0-2
|
GTGGAGCTCAT ACAA GAGGACAAGGCTTCAG +4bp
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
GTTCAGTATATAGTGGGTGGTGGAGCTCATACAAGAGGACAAGGCTTCAGCTTCGTCCAAACGC ( +4 bp，-0bp)
|
|
F0-2
|
CGCCACCTTGGCCATGGGGCGGTTCCCGACAAGGTTCTGGAGAGCCTGATGAAGTGGATC (+0bp，-10 bp)
|
|
Allele name and predicted product
|
998a
998b |
GTTCAGTATATAGTGGGTGGTGGAGCTCATACAAGAGGACAAGGCTTCAGCTTCGTCCAAACGC(47aa)turncate protein
|
| AACGCCACCTTGGCCATGGGGCGGTTCCCGAC..........AAGGTTCTGGA(77aa) turncate protein |
|
突变创建时间 |
2013-06-18
|
|
填表时间 |
2013/12/23
|
|
修改时间 |
2016/10/24
|
|
PI/负责人 (工作单位) |
|