Cellectis’ patented PulseAgile technology

In 2010, Cellectis acquired the assets of CytoPulse Sciences Inc., a Maryland-based company specializing in technology and equipment related to electroporation, which is a process using highly controlled electric fields that can be used to deliver messenger RNA (mRNA) or DNA molecules into cells. CytoPulse’s leading PulseAgile electroporation technology uses a unique electrical field wave-form that, in combination with a proprietary buffer solution, enables molecules, such as nucleases, to enter efficiently into the cell while maintaining high cell viability. PulseAgile uses a particularly effective combination of high voltage peaks, which are optimized to create transient holes in the cell membrane, followed by lower voltage pulses that help mRNA migrate into the cells. Critically, PulseAgile is optimized to preserve high cell viability and thus suited for large-scale manufacturing.

Importantly, PulseAgile provides a high-quality platform for delivering nuclease-encoding mRNA into T-cells, where it can be translated to generate an active nuclease protein that can access and specifically cut the cell’s genomic DNA. The mRNA molecules are rapidly degraded by the cell, which means that the nucleases are only expressed for a short time during which they operate the intended precise surgical DNA modifications.

Pulse parameters i.e. amplitude, duration, repetition frequency and shape affect electroporation efficacy

Exponential decay

Limited in efficacy and cell viability.

Rectangular wave

Second generation wave. Repeat wave increase payload.

Pulse Agile

Full optimized wave forms and frequency for better payload and viability.