February 20th, 2019, and Dr Julia Cricco (right) gathers her research team at the National University of Rosario (UNR), Argentina, to greet the new arrival; a small, grey box… This unremarkable-looking machine is a ‘Nucleofector’ from Lonza, purchased with Equipment Fund support from the Global Network for Neglected Tropical Diseases (NTDs). This device will transform the speed at which Julia’s group can progress towards identifying valid biological targets for developing new drugs against Chagas disease. The arrival of this one small box is therefore much celebrated, provoking excitement and big smiles all round!
The trypanosome parasites Leishmania (causing leishmaniasis) and Trypanosoma cruzi (causing Chagas disease) are ancient organisms with a unique biology. Their molecular mechanisms for operating essential processes, such as breathing and obtaining nutrients, are radically different from those used in cells from other species, including humans. This means that the genes controlling some of these processes in these parasites may be suitable as targets to use in screens for new drug compounds to treat both infections. However, this uniqueness also makes trypanosomes difficult to study; many standard genetic investigative techniques simply do not work in these ‘biologically odd’ cells.
Julia and her team have optimised CRISPR-Cas9 technology to remove and replace specific genes in T. cruzi. The procedure involves a transfection step, i.e. persuading the cell to take up the replacement DNA. The team in Rosario achieve this using electroporation – applying a pulse of electricity to briefly open pores in the parasite’s outer membrane and allow the replacement genetic material to enter the cell. However, uptake of the DNA is inefficient, typically taking weeks to achieve using their laboratory’s standard electroporation equipment, designed for use on simple bacterial cells. The ‘Nucleofector’ machine will enable these researchers to optimise their CRISPR process for T. cruzi, speeding up this step tenfold – something this team never dreamed would be possible. A reason for smiling indeed!
The NTD Network project is funded by the Global Challenges Research Fund (GCRF). The Equipment Fund is available to support purchases at UNR and other NTD Network member research organisations improve their infrastructure for research into Chagas disease and leishmaniasis. All NTD Network member research organisations in Asia and South America are eligible to apply. For further information, please get in touch; firstname.lastname@example.org.