Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (E. coli) | TOP10 | Invitrogen | C404010 | Cloning bacteria |
| Strain, strain background (E. coli) | BL21 (DE3) pLysS | Invitrogen | C606010 | Expression bacteria |
| Strain, strain background (E. coli) | MJF641 (Δ7) | Provided by Dr. Ian Booth (University of Aberdeen, UK) | Spheroplast strain | |
| Cell line (T. cruzi) | Y strain | ATCC | 50832 | Parasites used in this study |
| Cell line (Homo sapiens) | HFF | ATCC | SCRC-1041 CVCL_3285 |
Cells used to produce infective forms |
| Cell line (H. sapiens) | HEK293 | BEI resources | NR-9313 CVCL_0045 |
Cells used to produce infective forms |
| Transfected construct | Human T Cell Nucleofector Solution | LONZA | VPA-1002 | Transfection reagent for epimastigotes |
| Recombinant DNA reagent | pQE80L | QIAGEN | 32943 | Bacterial expression vector |
| Recombinant DNA reagent | Cas9/pTREXn | Addgene | RRID:Addgene_68708 | Vector for CRISPR-Cas9 -mediated KO |
| Recombinant DNA reagent | tdTomato/ pTREXb | Addgene | RRID:Addgene_68709 | Vector for KO donor cassettes |
| Recombinant DNA reagent | pUC_sgRNA | Addgene | RRID:Addgene_68710 | sgRNA backbone containing vector |
| Recombinant DNA reagent | pTREXn-eGFP | Addgene | RRID:Addgene_62544 | Expression vector for T. cruzi |
| Antibody | Anti-TcMscS | Cocalico Biologicals Inc | Generated for this work | Polyclonal antibody produced in guinea pig used for western blot and IFA analysis (1:1000 and 1:100, respectively). |
| Antibody | Anti-myc (mouse monoclonal) | SIGMA | RRID:AB_439694 (M4439) | Antibody used for western blot analysis and IFA (1: 5000 and 1:250, respectively) |
| Antibody | Anti-actin (mouse monoclonal) | SIGMA | RRID:AB_262137 (A3853) | Antibody used for IFA (1:250) |
| Antibody | Anti-Tubulin (mouse monoclonal) | SIGMA | RRID:AB_477579 (T5168) | Antibody used for IFA (1:500) and western blot (1:5000) |
| Antibody | Anti-trypanosome SSP1 (mouse monoclonal) | BEI Resources | NR-50891 | Used as trypomastigote marker for IFA (1:250) |
| Antibody | Anti-trypanosome SSP4 (mouse monoclonal) | BEI Resources | NR- 50892 | Used as amastigote marker for IFA (1:250) |
| Antibody | Anti- GFP (rabbit polyclonal) | Thermo Fisher | RRID:AB_221570 (A6455) | Antibody used for IFA (1:3000) |
| Chemical compound, drug | Fura2-AM | Thermo Fisher | F1221 | Reagent used for calcium measurements |
| Software, algorithm | pCLAMP11 | Molecular Devices | RRID:SCR_011323 | Electrophysiology acquisition and analysis software |
| Software, algorithm | Graph pad PRISM9 | http://www.graphpad.com | RRID:SCR_002798 | Statistical analysis and graph software |