作用点公知 細胞増殖抑制 生物活性未知
作用機構研究ツール
アッセイコンセプト確認
陽性化合物
創薬リード化合物 / 構造展開
用途特許出願
など、幅広い利用法
18.4 29.7 51.9
酵素阻害
膜蛋白質(受容体・
チャネル)
複製・転写・翻訳
シグナル伝達
細胞骨格・細胞周期
その他(RedOx)
30.0
25.0
20.0
15.0
10.0
5.0
0.0
100
-200
200
-300300
-400400
-500500
-600
600
-700
800
-900
1,000
-1,100
900
-1,000
700
-800
1,100
-1,200
1,200
-1,300
1,300
-1,400
1,400
-1,500>
1,500
O
O
H
H
O
OH
O
O
OH
O
H
H
OO
N
O
H
N
O
H
O
O
H
H
N
N
Br
Cl
O
O
O
O
O
Cl
Br
O
LIBRARY GENERATION
LIBRARY AMPLIFICATION
Reverse Transcription
(oligodT priming)
RNA Removal
Purification
PCR
Purification
Poly(A) RNA
Removal of RNA template
Random priming
cDNA library with adapters for Illumina sequencing
200 min 60 min
5’
5’
5’
3’
3’
3’
3’
5’
3’
3’
Tagged cDNA library
Double-stranded cDNA library
i7 index
(AAAAAAAA)
(AAAAAAAA)
(TTTTTTTTT)
5’
Second Strand
Synthesis
SEQUENCING - Read orientation for QuantSeq FWD
70 min 45 min
5’
i5 index (optional)
NBAAAAA.....AAAAAA
i7
index
Index 1 (i7)
Sequencing
Primer
Read 2 Read 2
Sequencing Primer
Read 1
Sequencing
Primer Read 1
(not recommended)
i5 index
(optional)
Index 2 (i5)
Sequencing
Primer
dCAS9dCAS9
dCAS9dCAS9
promoter
promoter Gene X
Gene X
KRAB
TA
sgRNA
sgRNA
NGG
NGG
CRISPRi and CRISPRa Variations
Activation-Fold
35
30
25
20
15
10
5
0
sg-control
MyoD sg-1
MyoD sg-2
MyoD sg-1+2
exp. #1 exp. #2 exp. #3
Catalog No. : T6234701-1
Cores : 90
Sixe : 2mm
Cut : 8μm
Format : 5×9
QA/QC : H&E
Slide 1
Slide 2
A
B
C
D
E
1 2 3 4 5 6 7 8 9
A
B
C
D
E
1 2 3 4 5 6 7 8 9
Cas9
sgRNA
TUNR
AAAAAAAAA
Your Gene of Interest
TUNR Gene of Interest
Introduction of the TUNR polyA track results
in reduced gene expression.
Wild Type
Short PolyA
Long PolyA
ProteinExpressionHighLow