Press Releases
CRISPR/Cas9 has transformed genome engineering by enabling precise and programmable modification of genomic DNA. Over the past decade, the technology has become an indispensable tool in biomedical research and has demonstrated significant potential for the treatment of cancer, genetic disorders, and hematological diseases.
Despite its broad applications, the success of CRISPR-based gene editing depends not only on the editing machinery itself, but also on efficient delivery to target cells. Delivering Cas9 and guide RNAs to the desired cell population while maintaining editing efficiency and minimizing off-target effects remains a major challenge for both research and therapeutic applications. As a result, the development of reliable delivery systems has become a central focus in the advancement of CRISPR technologies.
Among the available delivery approaches, lentiviral vectors are widely used because they efficiently transduce a broad range of cell types and support stable expression of CRISPR components. These properties have made lentiviral vectors valuable tools for gene knockout, gene regulation, pooled genetic screening, and disease modeling. At the same time, emerging delivery platforms such as virus-like particles (VLPs) and extracellular vesicles are being actively explored to enable transient CRISPR delivery and further improve safety profiles.
The following examples highlight recent advances in CRISPR-based research and illustrate how lentiviral and related delivery technologies are being applied to support genome editing studies and therapeutic development.
i-CRISPR: a personalized cancer therapy strategy through cutting cancer-specific mutations
Published in Molecular Cancer
Institution: Naval Medical University
The proposed i-CRISPR strategy designs and delivers a customized CRISPR-Cas9 system, namely cancer-targeted CRISPR scissors, which only cleaves cancer cell DNA according to cancer genome sequences and induces DNA breaks. Meanwhile, DNA damage repair inhibitors (DSBRi) are added to block the repair of broken DNA in cancer cells and trigger cell death. In the i-CRISPR system, the letter "i" represents DNA damage repair inhibitors, while "CRISPR" refers to the cancer-targeted editing tool. Since normal cells contain no cleavage sites for the designed CRISPR system, their DNA will not be cut or severely damaged. This strategy demonstrated selective targeting of cancer cells while minimizing damage to normal cells in preclinical models.
Research Models: HepG2 liver cancer cells, DU145 prostate cancer cells, organoids and PDX models
Pin Lyu et al., Nucleic Acids Res. 2019 Sep 26;47(17):e99.
Delivering Cas9/sgRNA ribonucleoprotein (RNP) by lentiviral capsid-based bionanoparticles for efficient ‘hit-and-run’ genome editing
Published in Nucleic Acids Research
Institution: College of Life Sciences, Anhui Normal University
Transient delivery of CRISPR components is an attractive strategy for reducing prolonged Cas9 expression, which may help minimize off-target editing and immune responses. To address this challenge, the authors developed a lentiviral capsid-based bionanoparticle system capable of packaging and delivering Cas9/sgRNA ribonucleoprotein (RNP) complexes. Using specific interactions between aptamers and aptamer-binding proteins (ABPs), the system efficiently assembled Cas9 RNPs into lentivirus-like particles (LVLPs) for transient genome editing.
The results show that replacing the Tetraloop of the sgRNA scaffold with a com aptamer does not affect sgRNA function. Com-modified sgRNA enables the assembly of Cas9/sgRNA RNP into lentivirus-like particles via specific interactions between ABP and aptamers, as well as between sgRNA and Cas9 protein. These RNP nanoparticles produce insertions and deletions at different target sites in various cell lines, with editing efficiency comparable to or higher than Cas9 mRNA-loaded LVLPs. The Cas9/sgRNA RNP system works rapidly with low off-target rates, enabling convenient and efficient delivery for transient Cas9 expression and high-efficiency genome editing.
Cell type: Human lymphoblastoid cells
Pin Lyu et al., Nucleic Acids Res. 2019 Sep 26;47(17):e99.
Systemic delivery of CRISPR/Cas9 to hepatic tumors for cancer treatment using altered tropism of lentiviral vector
Published in Biomaterials
Institution: Scripps Korea Antibody Institute
The lack of efficient in vivo delivery vectors has become a major obstacle for the therapeutic application of CRISPR/Cas9 nucleases. This study evaluated lentiviral vectors pseudotyped with hepatitis C virus (HCV) E1E2 envelope glycoproteins for in vivo delivery of CRISPR/Cas9 to liver tumors. The results indicate that E1E2-pseudotyped lentiviral vectors can selectively deliver Cas9 and sgRNA targeting kinesin spindle protein (KSP) to orthotopic Huh7 tumors in mice through specific binding to cellular receptors. This targeted delivery effectively disrupts the KSP gene and inhibits tumor growth. In addition, E1E2-pseudotyped lentiviral vectors are stable in human serum, highly cell-specific and induce low innate immune responses, making them promising delivery systems for CRISPR/Cas9-based cancer research and therapeutic development
Mouse strain: BALB/c nude mice (4-week-old, female)
Sungjin Lee et al, Biomaterials 2021 05;272.
Exosome-mediated delivery of Cas9 ribonucleoprotein complexes for tissue-specific gene therapy of liver diseases
Published in Science Advances
Institution: Zhejiang University
CRISPR-Cas9 is a powerful therapeutic technology. However, the lack of safe and efficient in vivo delivery systems, especially tissue-specific vectors, limits its wide clinical application. In vivo delivery of Cas9 ribonucleoprotein (RNP) has unique advantages, yet the large size of Cas9 RNP exceeds the loading capacity of most existing vectors. This study developed a novel genome editing delivery system named Exosome-RNP. Cas9 RNP is loaded into exosomes isolated and purified from hepatic stellate cells via electroporation. In vitro, Exosome-RNP achieves efficient cytoplasmic delivery of RNP. In vivo, it accumulates specifically in liver tissues. By targeting p53 upregulated modulator of apoptosis (PUMA), cyclin E1 (CcnE1), lysine acetyltransferase 5 (KAT5) and other molecules, Exosome-RNP exhibits strong therapeutic potential in mouse models of acute liver injury, chronic liver fibrosis and liver cancer. These findings highlight the potential of exosome-based delivery systems for tissue-specific CRISPR applications in liver diseases.
Research Models: Mouse models of acute liver injury, chronic liver fibrosis, and orthotopic hepatocellular carcinoma
Tao Wan et al., Sci Adv. 2022 Sep 16;8(37):eabp9435.
· Generation of monoclonal knockout cell lines
· Construction of full-length and customized CRISPR/Cas9 libraries
· Establishment of animal models (subcutaneous, orthotopic and intravenous injection models)
· Comprehensive project services for anti-tumor drug development
Service Process: Project consultation → Scheme design → Project breakdown → Project execution → Experimental report delivery
Related Experiments: High-throughput gene screening, target gene regulation, cell function verification, animal model construction, mechanism research
OBiO Technology is dedicated to the development and production of viral vectors. We provide multiple lentiviral packaging systems, including three-plasmid and four-plasmid systems. Our customized lentiviral products cover gene overexpression, gene interference, CRISPR/Cas9, Cumate-inducible expression system, Tet-on/off system and Cre-loxP system. We also supply ready-to-use lentiviruses for CRISPR/Cas9 knockout or activation libraries, single/dual autophagy labeling and luciferase reporter assays.
Our self-developed and patented Vpack lentiviral packaging system is designed to improve viral production efficiency and reduce the risk of failed packaging. It ensures more accurate lncRNA expression, brighter vector fluorescent signals and more stable virus production.
Request Now