| ZKO number |
ZKO582 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
zgc:158645
|
|
Ensembl ID |
ENSDARG00000060581
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
antisense strand
GGCTCAGAATCGGAGAAGATtgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 7
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
TAATAAGTTGGCGGTCCAGAAG
|
|
PCR primer (reverse) |
GTCCGTTCCCTAAGCAACAAGA
|
|
Length of expected PCR product (bp)
|
426
|
|
Length of expected RE digestion product (bp) |
n/a
|
|
Plasmids used for making Cas9 mRNA |
pGH-T7-zCAS9
|
| Plasmids?used?for?making?gRNA |
pT7-gRNA-ZKOpT7-gRNA-ZKO580
|
| Primers?used?for?making?gRNA |
Pr-ZKO 580
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
200
|
|
Dosage of injected gRNA (ng/ul)
|
25
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
90
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
1 dpf
|
|
Mutation efficiency in F0 embryos (%)
|
36
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
50% (2/4)
|
|
Wild-type (WT) sequence
|
AGATGTGGTTCAGTGATTCTGGAGCCAATCTTCTCCGATTCTGAGCCCACGGCAGAGGCCCCCCTGGGGATGGGGGCTACTT
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
AGATGTGGTTCAGTGATTCTGGAGCCAATC---......---CAGAGGCCCCCCTGGGGATGGGGGCTACTT(22bp deletion)
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
AGATGTGGTTCAGTGATTCTGGAGCCAATC---......---CAGAGGCCCCCCTGGGGATGGGGGCTACTT(22bp deletion)
|
|
F0-2
|
|
|
Allele name and predicted product
|
582a
582b |
AGATGTGGTTCAGTGATTCTGGAGCCAATC---......---CAGAGGCCCCCCTGGGGATGGGGGCTACTT(22bp deletion) Truncated protein (267aa)
|
| |
|
突变创建时间 |
2014-04-10
|
|
填表时间 |
2013/12/18
|
|
修改时间 |
2015/5/29
|
|
PI/负责人 (工作单位) |
舒晓东(广州生物医药与健康研究院)
|
