Case Study: Synthetic sgRNAs Enable Researchers to Study Viral Infection in Resting Human CD4+ T Cells
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Case Study: Synthetic sgRNAs Enable Researchers to Study Viral Infection in Resting Human CD4+ T Cells
In a new case study, Synthego and Lonza discuss their recent collaboration with Dr. Manuel Albanese and exciting results in gene editing of resting human CD4+ T cells. This case study highlights how to achieve efficient gene editing in CD4+ T cells, overcoming the traditional challenges related to cell viability and maintaining resting state. The corresponding protocol provides a step-by-step guideline along with a tips and tricks section for performing gene editing using Synthego’s sgRNA in combination with Lonza's 4D-Nucleofector® System.
By implementing an optimized workflow that includes interleukin supplementation, sgRNA, and the 4D-Nucleofector® System, Dr. Manuel Albanese and his colleagues achieved knock out rates greater than 98% of up to six genes simultaneously in resting CD4+ T cells.
The advancements presented in this case study further illuminate the potential of genome editing in hard-to-transfect primary cells to drive research in human immunology and viral infection mechanisms.
Based on Albanese et al., Nature Methods 19, 81-89 (2022)
Ready to CRISPR your cells of interest? Register to download a complimentary copy of the protocol
Access the detailed protocol of Synthego and Lonza that outlines the workflow of delivering ribonucleoprotein (RNP) complexes into human primary resting CD4+ T cells. Using the Lonza 4D-Nucleofector® System and Synthego’s chemically modified sgRNA, this protocol, successfully employed by Dr. Albanese, provides a robust framework for advancing your own research in gene editing of resting CD4+ T cells. Of note, further recommendations from Synthego and Lonza on how to adapt the protocol to other cell types and higher throughputs (e.g. 96-well format) are included in a tips and tricks section.
Please fill in the form to get your copy of the protocol and related information.
