一、服務介紹
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圖一:CRISPR library 使用方法示意圖,以已經建構完成的 CRISPR library (質體),製作慢病毒 library,感染細胞並經過篩選後,找出 phenotype 改變的細胞株,進而發現相關的基因。 |
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圖二:全客製化 CRIPSR Library,針對有興趣的疾病或路徑,設計出專屬的 CRISPR Library。(1) 半導體晶片方式合成 Oligo:正確性和均勻度更高、交期更快。(2) 一步就位:直接接到客戶載體上,省下 Cloning 時間。(3) NGS 檢測,>99% 覆蓋率。 |
二、服務應用
針對目標基因一次做大量的篩選:
可以一次針對所有基因做 knock-out 或是促進表現,也可以選購 Pathway-focused CRISPR gRNA library,針對特定訊息路徑或疾病,一次檢測所有相關基因。
三、服務方案
針分三種 Library,可一次針對所有基因做 knockout (表一) 或促進表現 (表三),或是針對特定疾病或訊息傳遞路徑 (表二)。
表一:GeCKO library for genome-scale knockout:一次針對所有基因 Knockout
Library 名稱 | 序列資訊 | 貨號 | 載體 | 包裝 |
Human GeCKO Library A | 65,383 sequences: 3 sgRNA targeting each of 19,050 genes; 4 sgRNA targeting each of 1,864 miRNA; 1,000 control (non-targeting) sgRNAs. |
SC1771 | pLentiCRISPR v2 | 25 ug Mixed pool plasmid (所有 gRNA 質體混合成一管) |
SC1774 | pLentiGuide-Puro, pLentiCas9-Blast | |||
Human GeCKO Library B | 58,028 sequences: 3 sgRNA targeting each of 19,050 genes; 1,000 control (non-targeting) sgRNAs. |
SC1777 | pLentiCRISPR v2 | |
SC1780 | pLentiGuide-Puro, pLentiCas9-Blast | |||
Mouse GeCKO Library | 67,405 sequences: 3 sgRNA targeting each of 20,611 genes; 4 sgRNA targeting each of 1,175 miRNA; 1,000 control (non-targeting) sgRNAs |
SC1783 | pLentiCRISPR v2 | |
SC1843 | pLentiGuide-Puro, pLentiCas9-Blast | |||
Mouse GeCKO Library B | 62,804 sequences: 3 sgRNA targeting each of 20,611 genes; 1,000 control (non-targeting) sgRNAs. |
SC1849 | pLentiGuide-Puro, pLentiCas9-Blast |
表二:Pathway-focused CRISPR gRNA library:針對特定訊息路徑或疾病,一次檢測所有相關基因
針對訊息路徑或疾病 | 基因數目 | gRNA 數目 |
ABC Transporter | 100 | 298 |
Acute myeloid leukemia | 45 | 135 |
Adipogenesis | 126 | 378 |
Adult Stem Cells | 58 | 174 |
Alzheimer's disease | 94 | 281 |
Angiogenesis | 92 | 276 |
Apoptosis | 68 | 204 |
B cell receptor signaling pathway | 58 | 174 |
Bladder cancer | 37 | 111 |
cAMP & calcium Signaling Pathway | 91 | 271 |
Cardiovascular Disease | 91 | 273 |
Cell Cycle | 84 | 252 |
Chemokine signaling pathway | 134 | 402 |
Chronic myeloid leukemia | 57 | 171 |
Colorectal cancer | 54 | 162 |
Cytochrome P450 | 57 | 171 |
Diabetes | 44 | 132 |
Drug Metabolism | 34 | 102 |
Drug Resistance | 348 | 1044 |
Drug transporters | 84 | 252 |
ES Cell Differentiation | 334 | 1002 |
Extracellular Matrix & Adhesion | 84 | 252 |
GPCR | 280 | 840 |
Growth Factor | 161 | 483 |
Histone Modification | 259 | 776 |
Hormone Activity | 108 | 324 |
Ion Channels (Potassium) | 67 | 201 |
Nuclear Hormone Receptors | 117 | 351 |
Oncogenes | 289 | 867 |
Tumor Metastasis | 60 | 180 |
Tumor Suppressor | 720 | 2155 |
WNT Pathway | 92 | 276 |
表三:CRISPR transcriptional activation SAM library:一次針對所有基因促進表現
Library 名稱 | 序列資訊 | 貨號 | 載體 | 包裝 |
Human SAM Library | 70,781 gRNA sequences: 3 sgRNA targeting each of 23,430 unique coding isoforms, 491 non target control |
SC1770 | pLenti sgRNA(MS2)_zeo; pLenti dCas9-VP64_Blast; pLenti MS2-P65-HSF1-Hygro. |
25 ug Mixed pool plasmid (所有 gRNA 質體混合成一管) |
SC1754 | pLenti sgRNA(MS2)_puro; pLenti dCas9-VP64_Blast; pLenti MS2-P65-HSF1-Hygro. |
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Mouse SAM Library | 69,716 gRNA sequences: 3 sgRNA targeting each of 23,075 unique coding isoforms, 491 non target control |
SC1844 | pLenti sgRNA(MS2)_puro; pLenti dCas9-VP64_Blast; pLenti MS2-P65-HSF1-Hygro. |