Accelerate research, empower industry
CRISPR Library Screening Services
Hysigen leverages its proprietary CRISPRSeek™ library screening platform and state-of-the-art algorithms to deliver three customizable, high-impact library screening services: CRISPR knockout (CRISPR-KO), CRISPR activation (CRISPRa), and CRISPR inhibition (CRISPRi). Our seamless solutions encompass the entire workflow, from sgRNA design to viral packaging, cell transfection, high-throughput sequencing, and in-depth data analysis. Comprehensive solutions empower groundbreaking research on gene function and drug target discovery. By leveraging efficient, precise, and intelligent technical platforms, we are dedicated to fulfilling diverse scientific research aspirations.
CRISPR KO Pooled Libraries An efficient gene knockout is achieveded by inducing frameshift mutations in target gene exons.
Application: Enable genome-wide loss-of-function genetic screening to identify disease-associated genes. By leveraging CRISPR-KO library screening, researchers can precisely localize crucial targets among thousands of genes, laying the groundwork for the discovery of novel therapeutic targets. | CRISPRa Pooled Libraries CRISPR activation is facilitated by fusing dCas9 with VP64, thereby upregulating target gene expression.
Application: Enable gain-of-function genetic screening to study how gene overexpression affects phenotypes. By using CRISPRa (CRISPR activation) library to activate specific genes, researchers can identify potential candidates whose activation can improve cell survival rates. This approach can reveal promising new gene targets for further exploration. | CRISPRi Pooled Libraries Gene silencing is achieved by binding dCas9-VP64 to taget DNA sequence, therby blocking the target gene expression.
Application: Enable research on the functions of essential genes and investigation of the regulatory mechanisms of non-coding RNAs. CRISPR interference (CRISPRi) library can be used to screen downstream regulatory factors associated with crucial genes in cancer cells, thereby uncovering novel signaling pathways and developing new strategies for inhibiting tumor growth. |
1. Library Construction: We provide custom guide RNA (gRNA) design, primer synthesis, and vector construction, supporting library size ranging from 1,000 to 92,000.
2. Library Amplification: We ensure 99% coverage and maintain uniformity with less than 10% variability.
3. Pooled Library Virus Packaging: We produce high-titer library virus particles available in specifications ranging from 1 to 100 mL.
4. Pooled Library Cells: Our automated production can provide over 400 cell types, achieving more than 95% coverage of pooled library cells.
5. Functional Screening: We provide comprehensive screening protocols, with optional cell flow cytometry sorting platforms.
6. Data Analysis: Our deep sequencing and data interpretation utilize the CRISPRSeek™ algorithm platform, delivering rich data, including charts and figures.
| Service Modules | Descriptions | Deliverables |
| Custom sgRNA library design, amplification, and quality control | sgRNA design, synthesis of sgRNA-oligo pool, construction of pooled library plasmids, amplification of library plasmidspooled , and NGS-based quality control | Pooled library plasmids: 100 μg, along with NGS-based report (coverage > 99%) |
Custom Cas9 stable cell lines | Cas9 lentivirus packaging, construction of Cas9 stable cell lines, and Editing-activity validation | Cas9 stable cell lines (1×10⁶ cells/vial), 2 vials |
Pooled library lentivirus packaging | Packaging and titer determination of pooled library lentivirus (cell titration method), ensuring functional lentiviral titer above 1 x 10⁸ TU/mL | 1 mL: Total of 1×10⁸ TU 5 mL: Total of 5×10⁸ TU 10 mL: Total of 1×10⁹ TU Viral titer test reports. |
Pooled library cell construction | Optimization of transfection conditions with MOI < 0.3, and construction of pooled library cells | Small scale: total of 1×10⁷ cells Medium scale: total of 5×10⁷ cells Large scale: total of 1×10⁸ cells |
Functional screening assays | Custom functional screening services: drug screening, viral infection screening, bacterial infection screening, flow cytometry sorting, etc. | Experimental reports |
NGS analysis | sgRNA sequencing library construction and NGS sequencing, bioinformatics analysis | Experimental report, raw NGS data |
| Typical Applications
Genome-wide screening: Using CRISPR knockout (CRISPR-KO) libraries to perform genome-wide screens in cancer cells, aiming to identify genes associated with drug resistance.
Signaling pathway research: Using CRISPR interference (CRISPRi) libraries to target signaling pathway genes, which helps to elucidate the functions and regulatory roles of these genes.
Drug target discovery: Using CRISPR activation (CRISPRa) libraries to upregulate gene expression in cells, allowing for the identification of novel genes that enhance specific phenotypes.
| Typical Application Scenarios of CRISPRSeek™ Library Screening Services:
1. Application of CRISPR-KO Library Screening in Genome-Wide Genetic Screening
Case Study: Identification of host factors essential for SARS-CoV-2 infection. Researchers used CRISPR-KO libraries to identify key host genes critical for viral entry. They discovered that knocking out ACE2 and TMPRSS2 significantly reduced viral infection efficiency.
Reference:
Daniloski, Z., et al. (2021). "Identification of Required Host Factors for SARS-CoV-2 Infection in Human Cells." Cell, 184(1), 92-105.e16. https://doi.org/10.1016/j.cell.2020.10.030
2. Application of CRISPRa Library Screening in Drug Target Discovery
Description: Screening target genes, which are critical to cell survival or disease progression, to identify potential drug targets.
Case study: Studies on BRAF inhibitor resistance. CRISPRa library screening revealed that activation of EGFR leads to the resistance to BRAF inhibitors, providing a novel strategy for combination therapies.
Reference:
Konermann, S., et al. (2015). "Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex." Nature, 517(7536), 583-588. https://doi.org/10.1038/nature14136
3. Application of CRISPRi Library Screening in Essential Gene Research
Description: Investigating gene functions by knocking down essential genes using CRISPRi.
Case Study: For the first time, CRISPR interference (CRISPRi) libraries were used to knock down genes throughout the entire genome, including essential genes. This approach was combined with CRISPR activation (CRISPRa) to investigate gene expression regulation.
Reference: Gilbert et al. (2014) "Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation." Cell. 2014 Oct 23;159(3):647-61. https://doi.org/10.1016/j.cell.2014.09.029
Hysigen provides comprehensive CRISPRSeekTM library screening services, supporting a wide variety of applications. These include genome-wide screening, targeted gene family analysis, signaling pathway exploration, drug target discovery, non-coding RNA research, essential gene studies, and epigenetic regulation. Our services have shown significant potential in areas such as cancer research, neuroscience, regenerative medicine, and industrial biotechnology. Contact us today to start your journey toward precise gene screening!
Why choose Hysigen Biosciences? At Hysigen Biosciences, we prioritize innovative technology, efficient execution, and reliable outcomes as our core values. We provide professional support to researchers around the globe. Whether you need assistance with whole-genome screening or research on specific pathways, we can tailor solutions to help you overcome challenges in your scientific research.
|
|
![](_script:Site.hoverQrCode("//12794934.s61i.faiusr.com/4/AD0ItviMBhAEGAAgs6DslQYo1MGy9wcw0gE40gE.png",this);)
![](_script:Site.hoverQrCode("//12794934.s61i.faiusr.com/4/AD0ItviMBhAEGAAgouDulQYo7IavQzDIATjIAQ.png",this);)