A Sponge Can Simplify Lentivirus Transduction

Lentiviral transduction is a popular delivery platform for applications such as CAR-T cell engineering or CRISPR-Cas9 gene editing, as it can transduce both dividing and non-dividing cells and accommodate large payloads.¹ However, learning how to work with lentiviruses can be tricky. Researchers can “spend significant time troubleshooting and optimizing each step in the process—wasting months of work and lab resources,” said Thomas Quinn, a virologist working for Takara Bio.

In the standard lentiviral transduction protocol, cells sit at the bottom of a well while payload-containing virus is in solution on top. In this configuration, most of the virus never contacts the cells. Attempts to increase virus-cell contact using mechanical centrifugation (spinoculation) or chemical enhancement offer only limited gains in efficiency and can exert unpredictable downstream effects on the cells.

By contrast, microfluidics places cells and viruses together within narrow spaces to increase transduction efficiency. Unfortunately, these approaches require expensive specialized equipment. Considering this, Quinn and his colleagues at Takara Bio developed the Lenti-X™ Transduction Sponge, an alginate sponge that mimics microfluidic approaches by co-localizing cells and virus within its 20–300 µm pores. The sponge is easy to use: just add cells and viruses, incubate for 24 hours, and elute with buffer to release healthy, transduced cells ready for downstream use.

Quinn posited that the biggest advantage of the Lenti-X™ Transduction Sponge is its flexibility and accessibility. The sponge is cell- and particle-type agnostic, and it works with difficult-to-transduce cells including hematopoietic stem cells, natural killer cells, and T cells, as well as other viral types such as retroviruses and adenoviruses. “Any researcher, whether a brand-new graduate student or seasoned lentivirus expert, can achieve successful transductions using microfluidics,” said Quinn.

Read more about simplifying the lentivirus production and transduction workflow.