Gene editing of putative cAMP and Ca²⁺ -regulated proteins using an efficient cloning-free CRISPR/Cas9 system in Trypanosoma cruzi.

ABSTRACT: Trypanosoma cruzi, the agent of Chagas disease, must adapt to a diversity of environmental conditions that it faces during its life cycle. The adaptation to these changes is mediated by signaling pathways that coordinate the cellular responses to the new environmental settings. Cyclic AMP (cAMP) and Calcium (Ca²⁺ ) signaling pathways regulate critical cellular processes in this parasite, such as differentiation, osmoregulation, host cell invasion and cell bioenergetics. Although the use of CRISPR/Cas9 technology prompted reverse genetics approaches for functional analysis in T. cruzi, it is still necessary to expand the toolbox for genome editing in this parasite, as for example to perform multigene analysis. Here we used an efficient T7RNAP/Cas9 strategy to tag and delete three

SUBMITTER: Chiurillo MA 

PROVIDER: S-EPMC10841170 | biostudies-literature | 2023 Nov-Dec

REPOSITORIES: biostudies-literature