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. 2020 Aug 13;10:408. doi: 10.3389/fcimb.2020.00408

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Copyright © 2020 Jesus-Santos, Lobo-Silva, Ramos, Descoteaux, Lima, Borges and Farias.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Generation of LPG2 knockout using CRISPR/Cas9. (A) Western blot analysis of L. infantum promastigotes expressing Cas9 (gRNA440 and gRNA516). (B) Growth curves of L. infantum wild-type (WT) and L. infantum-Cas9 (gRNA440 and gRNA516) promastigotes. (C) Agglutination assay using the CA7AE monoclonal antibody and associated growth curves. (D) Agglutination assay using Ricin 120 lectin and associated growth curves, demonstrating the selection of Δlpg2. The arrows indicate the time points where antibody (CA7AE) or lectin (Ricin-120) were added to the cultures, exemplifying the typical agglutination results observed.