Synthetic sgRNAs Enable Researchers To Study Viral Infection In Resting Humans CD4+ T Cells

Genome editing in resting CD4+ T cells presents significant challenges, including limited viability and low editing efficiency, which historically constrain downstream functional assays such as pathway analysis. In a groundbreaking case study conducted at the Max von Pettenkofer Institute and Gene Center in Munich, Germany, Dr. Manuel Albanese and Dr. Adrian Ruhle, under the guidance of Dr. Oliver Keppler, demonstrated how these challenges could be overcome.

By leveraging a combination of optimized cell culture conditions, Synthego’s synthetic sgRNA, and Lonza’s 4D-Nucleofector® technology, the team achieved unprecedented knockout editing efficiencies while maintaining high viability in primary T cells. The study highlights their success in achieving high and consistent knockout efficiencies in resting primary human CD4+ T cells, enabling experiments involving multigene knockouts. Furthermore, sustained cell viability allowed for robust downstream functional studies, providing a valuable framework for advancing research in primary T cell biology and genome editing.