CRISPR遺伝子転写抑制(CRISPRi)により、研究者はDNAを編集することなく、転写をブロックすることによって特定の遺伝子機能をダウンレギュレートすることができます。詳細については、CRISPRiアプリケーションのページをご覧ください。.
Horizon CRISPRmod CRISPRiシステムには、遺伝子特異的なCRISPRiガイドRNAと転写リプレッサードメイン(SALL1およびSDS3)に融合し触媒活性が不活化されたCas9(dCas9)の2つのコンポーネントが必要です。当社のCRISPRi試薬は、遺伝子転写抑制を介して遺伝子の機能を研究するための簡便で効率的なツールセットを提供します。
CRISPRi試薬の選択
CRISPRiの実験条件には多くのオプションと考慮事項があります。長時間のアッセイ(120時間以上)には、レンチウイルスsgRNAをお勧めします。短時間のアッセイでは、化学合成sgRNAは、通常、ガイドRNA発現ベクターよりもより強力な遺伝子転写抑制を提供します。
CRISPRi dCas9-SALL1-SDS3ソース | ガイドRNAフォーマット | デリバリー方法 | メリットと推奨される使い方 |
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CRISPRi dCas9-SALL1-SDS3 lentiviral particle |
CRISPRi synthetic sgRNA(化学合成sgRNA) | トランスフェクションまたはエレクトロポレーション |
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CRISPRi lentiviral sgRNA | 形質導入 |
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CRISPRi dCas9-SALL1-SDS3 mRNA | CRISPRi synthetic sgRNA(化学合成sgRNA) | コトランスフェクションまたはエレクトロポレーション |
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Order CRISPRi guide RNA
CRISPRi synthetic sgRNA
The most rapid CRISPRi system, with repression within 24 hours post-transfection.
CRISPRi lentiviral sgRNA
An ideal delivery method for CRISPRi in difficult-to-transfect cells, or for longer term repression.
CRISPRi dCas9 solutions
dCas9-SALL1-SDS3 mRNA
Co-transfect with synthetic sgRNA for transient CRISPRi in lentiviral-free workflows.
dCas9-SALL1-SDS3 lentiviral particles
Create a stable dCas9 expressing cell line, ready for CRISPRi studies.
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CRISPRi gene repression is observed 24 hours post-transfection and is maximal 48-72 hours post transfection
CRISPRi synthetic guide RNA
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CRISPRi synthetic sgRNA
The most rapid CRISPRi system, with repression within 24 hours post-transfection. Synthetic RNAs are modified for nuclease resistance and available as pooled or individual reagents.
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CRISPRi synthetic sgRNA positive controls
Control sgRNAs targeting well-characterized genes to determine the effectiveness of your experimental conditions
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CRISPRi synthetic sgRNA non-targeting controls
Non-targeting controls to evaluate baseline cellular responses to CRISPRi components in the absence of target-specific crRNA
CRISPRi lentiviral guide RNA
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CRISPRi lentiviral sgRNA
Lentiviral particles of vector-based single guide RNA are predesigned for genome-wide coverage of human genes
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CRISPRi lentiviral sgRNA positive controls
Control sgRNAs targeting well-characterized genes to determine the effectiveness of your experimental conditions
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CRISPRi lentiviral sgRNA non-targeting controls
Non-targeting controls to evaluate baseline cellular responses to CRISPRi components in the absence of target-specific sgRNA
CRISPRi dCas9-SALL1-SDS3
Nuclease-deactivated Cas9, fused to our proprietary transcriptional repressor domains (SALL1 and SDS3).
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CRISPRi dCas9-SALL1-SDS3 mRNA
Co-transfect with synthetic sgRNA using Dharmafect transfection reagents for transient dCas9-SALL1-SDS3 expression in lentiviral-free workflows.
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CRISPRi dCas9-SALL1-SDS3 lentiviral particles
Create a stable dCas9-SALL1-SDS3 expressing cell line, ready for CRISPRi studies.
CRISPRi screening libraries
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Custom cherry-pick libraries
Upload your own gene list to customize and order plates of predesigned sgRNA for CRISPRi studies across tens or thousands of genes
Horizon's CRISPRi workflows
We offer multiple workflow options when using CRISPRi for transcriptional repression
- Transduce with dCas9-SALL1-SDS3 lentiviral particles to generate CRISPRi-ready, stable dCas9-SALL1-SDS3 expressing cells. Then introduce either A) synthetic or B) lentiviral sgRNA to knockdown your target gene(s). This system is ideal for screening, extended time point assays or when working with difficult-to-transfect cell types.
- C) Co-transfect or electroporate dCas9-SALL1-SDS3 mRNA with synthetic sgRNA, then enrich cell populations using fluorescence or puromycin resistance options. This system is best suited for rapid, transient gene repression studies.
CRISPRi workflows