Article
Microbiology
Gabriela de A. Burle-Caldas, Nailma S. A. dos Santos, Julia T. de Castro, Fernanda L. B. Mugge, Viviane Grazielle-Silva, Antonio Edson R. Oliveira, Milton C. A. Pereira, Joo Luis Reis-Cunha, Anderson Coqueiro dos Santos, Dawidson Assis Gomes, Daniella C. Bartholomeu, Nilmar S. Moretti, Sergio Schenkman, Ricardo T. Gazzinelli, Santuza M. R. Teixeira
Summary: Trans-sialidases (TS) are enzymes present on the surface of Trypanosoma cruzi, the causative agent of Chagas disease, and play a crucial role in the late stages of intracellular development and parasite egress. In this study, TS knockout parasites were generated using CRISPR-Cas9 technology, resulting in impaired parasite egress from infected cells. These TS mutants lost their ability to cause infection in vivo but provided full protection against a challenge infection with a virulent strain, indicating their potential as a live attenuated vaccine against Chagas disease.
Article
Microbiology
Gabriela de A. Burle-Caldas, Nailma S. A. dos Santos, Julia T. de Castro, Fernanda L. B. Mugge, Viviane Grazielle-Silva, Antonio Edson R. Oliveira, Milton C. A. Pereira, Joao Luis Reis-Cunha, Anderson Coqueiro dos Santos, Dawidson Assis Gomes, Daniella C. Bartholomeu, Nilmar S. Moretti, Sergio Schenkman, Ricardo T. Gazzinelli, Santuza M. R. Teixeira
Summary: Trans-sialidases play a crucial role in the virulence of Trypanosoma cruzi, and using CRISPR-Cas9, aTS mutant parasites were generated which lost infectivity in vivo but provided full protection against a challenge infection with a virulent strain.
Article
Immunology
Jose L. Saenz-Garcia, Beatriz S. Borges, Normanda Souza-Melo, Luiz V. Machado, Juliana S. Miranda, Lisandro Alfonso Pacheco-Lugo, Nilmar S. Moretti, Richard Wheleer, Lia C. Soares Medeiros, Wanderson D. DaRocha
Summary: The flagellum of Trypanosomatids contributes to multiple functions, and this study explores the role of Trypanin in T. cruzi. The deletion of Trypanin affects the growth and motility of T. cruzi epimastigotes, as well as their infection capacity.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2022)
Article
Fisheries
Qian Li, Hong Yu, Qi Li
Summary: Genome editing using CRISPR/Cas9 methodology holds great potential for improving aquaculture, but the screening and identification of successfully edited individuals remains challenging. In this study, we developed and optimized a high-resolution melting analysis (HRM) assay to rapidly screen and identify CRISPR/Cas9-edited oysters. The HRM assay showed sensitivity in detecting temperature shifts and successfully identified oysters with indel mutations. This reproducible, sensitive, and efficient method has the potential to accelerate genetic improvement in aquaculture.
Review
Entomology
Thu N. M. Nguyen, Amanda Choo, Simon W. Baxter
Summary: The sterile insect technique (SIT) is a pest control strategy that involves mass-rearing, sterilization, and release of male insects to reduce the population of insects that pose threats to agriculture or human health. Developing genetic sexing strains with conditional temperature sensitive lethal mutations offers potential to eliminate female embryos through heat treatment. The challenge remains in developing robust sex-separation systems for SIT programs to improve efficiency and cost-effectiveness.
Article
Multidisciplinary Sciences
Yu Fu, Zhenglin Zhu, Geng Meng, Rijun Zhang, Yueping Zhang
Summary: Post-translational modifications of histone proteins play a crucial role in gene expression and cell fate decisions. Developing efficient methods for introducing mutations to histones is vital for studying these modifications. By utilizing a CRISPR-Cas9 based system in yeast cells, researchers successfully generated multiple histone mutants, presenting a valuable tool for the field of epigenetics.
SCIENTIFIC REPORTS
(2021)
Article
Multidisciplinary Sciences
Arvind Sharma, Michael N. Pham, Jeremiah B. Reyes, Randeep Chana, Won C. Yim, Chan C. Heu, Donghun Kim, Duverney Chaverra-Rodriguez, Jason L. Rasgon, Robert A. Harrell, Andrew B. Nuss, Monika Gulia-Nuss
Summary: Ticks have the ability to transmit various debilitating pathogens, but research on ticks has been lacking compared to other arthropod vectors due to challenges in applying genetic and molecular tools. This study reports a successful embryo injection protocol for the black-legged tick and demonstrates the use of CRISPR-Cas9 for genome editing. The results provide innovative tools for tick research.
Article
Immunology
Thaise Lara Teixeira, Miguel Angel Chiurillo, Noelia Lander, Cassiano Costa Rodrigues, Thiago Souza Onofre, Eden Ramalho Ferreira, Camila Miyagui Yonamine, Julia de Gouveia Santos, Renato Arruda Mortara, Claudio Vieira da Silva, Jose Franco da Silveira
Summary: P21 is an immunomodulatory protein in Trypanosoma cruzi that plays crucial roles in host cell invasion, internalization, and amastigote proliferation.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2022)
Article
Plant Sciences
Bicko Steve Juma, Asunta Mukami, Cecilia Mweu, Mathew Piero Ngugi, Wilton Mbinda
Summary: This study successfully developed cassava plants with lower levels of cyanogenic glycoside and cyanide content through CRISPR/Cas9-mediated targeted mutagenesis. This provides an alternative approach for breeding non-toxic cassava varieties.
FRONTIERS IN PLANT SCIENCE
(2022)
Review
Biochemistry & Molecular Biology
Caixia Gao
Summary: Genome editing has the potential to revolutionize plant breeding and address the challenge of feeding a growing population in the face of climate change. Newly developed techniques and strategies offer significant opportunities for plant improvement and crop production, but challenges must be overcome to realize the full potential of this technology.
Article
Biochemistry & Molecular Biology
Ramona Gruetzner, Patrick Martin, Claudia Horn, Samuel Mortensen, Erin J. Cram, Carolyn W. T. Lee-Parsons, Johannes Stuttmann, Sylvestre Marillonnet
Summary: The recent discovery of the mode of action of the CRISPR/Cas9 system has provided biologists with a useful tool for generating site-specific mutations in genes of interest. In this study, optimizing the efficiency of the Cas9 nuclease in generating mutations in target genes by investigating its nucleotide and/or amino acid sequence features led to the finding that adding introns to the Cas9 coding sequence significantly improved editing efficiency. The intronized Cas9 gene was shown to be effective in generating mutant phenotypes in various plants, highlighting the importance of sequence modifications for enhancing CRISPR/Cas9 genome editing.
PLANT COMMUNICATIONS
(2021)
Article
Plant Sciences
Chong Ren, Yanfei Liu, Yuchen Guo, Wei Duan, Peige Fan, Shaohua Li, Zhenchang Liang
Summary: This study identified multiple effective promoters in grapevine and achieved multiplex genome editing by optimizing the CRISPR/Cas9 system, leading to increased editing efficiency. Simultaneous editing of TMT1 and TMT2 genes resulted in reduced sugar levels in grapes, highlighting the role of these genes in sugar accumulation. The findings provide valuable materials for the optimization of CRISPR/Cas9 system and expand the toolbox of grape genome editing for basic research and molecular breeding.
HORTICULTURE RESEARCH
(2021)
Article
Plant Sciences
Mengxi Bi, Zhijun Wang, Keyan Cheng, Yiqing Cui, Yi He, Jian Ma, Mingfang Qi
Summary: This study successfully applied the CRISPR/Cas9 large-scale screening strategy in tomato and constructed a CRISPR/Cas9 mutant library. The study found that homozygous and biallelic mutations occurred at higher frequencies in the mutant population. Additionally, a small-scale CRISPR/Cas9 library enhanced the efficacy of single sgRNA recognition in positive plants.
PLANT PHYSIOLOGY AND BIOCHEMISTRY
(2023)
Article
Cell Biology
Mayara S. Bertolini, Roberto Docampo
Summary: In this study, we generated TcMICU1/TcMICU2 double knockout cell lines using CRISPR/Cas9 genome editing and found that these knockout cell lines were unable to uptake calcium at low extramitochondrial calcium concentrations. Further investigation revealed that Mg2+ negatively regulates the mitochondrial calcium uptake mediated by MCUC.
Article
Plant Sciences
Bhuvan Pathak, Chandan Maurya, Maria C. Faria, Zahra Alizada, Soumen Nandy, Shan Zhao, Muhammed K. Jamsheer, Vibha Srivastava
Summary: This study demonstrates the efficiency of CRISPR/Cas9 in creating mutations in essential genes in rice, although the mutagenesis rates vary among different genes. Additionally, the study shows that the generated mutants exhibit phenotypes such as sterility or early senescence.
Article
Biochemistry & Molecular Biology
Noelia Lander, Miguel A. Chiurillo, Roberto Docampo
Summary: Trypanosoma cruzi is a unicellular parasite responsible for Chagas disease, with a digenetic life cycle alternating between mammalian and insect hosts. This parasite must adapt to various environmental conditions such as temperature, nutrient availability, pH, and oxidative stress, which are regulated by signaling pathways. This review will focus on the signaling pathways and second messengers involved in the responses to these environmental changes.
MOLECULAR MICROBIOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Brian S. Mantilla, Leticia D. Do Amaral, Henning J. Jessen, Roberto Docampo
Summary: The study highlights the importance of highly phosphorylated IPs in the life cycle of T. cruzi, identifying new highly phosphorylated IPs and the kinases involved in their synthesis.
ACS CHEMICAL BIOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Brian S. Mantilla, Karunakaran Kalesh, Nathaniel W. Brown, Dorothea Fiedler, Roberto Docampo
Summary: This study identified potential targets of 5-IP7 in Trypanosoma cruzi using pull-down experiments and mass spectrometry analysis, revealing new functional roles of 5-IP7 in this divergent eukaryote and providing potential new targets for chemotherapy.
MOLECULAR MICROBIOLOGY
(2021)
Review
Biochemistry & Molecular Biology
Roberto Docampo, Guozhong Huang
Summary: Trypanosoma cruzi and the T. brucei group of parasites are neglected diseases affecting millions of people globally. These microorganisms have complex life cycles involving insect vectors and mammalian hosts, and possess unique signaling pathways and receptor adaptations that differentiate them from their mammalian counterparts.
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH
(2021)
Article
Microbiology
Raquel S. Negreiros, Noelia Lander, Miguel A. Chiurillo, Anibal E. Vercesi, Roberto Docampo
Summary: Pyruvate, the end product of glycolysis, is transported into the mitochondria through the mitochondrial pyruvate carriers MPC1 and MPC2. Knockout of MPC1 and MPC2 in Trypanosoma cruzi affects pyruvate-driven respiration, host cell invasion, and intracellular replication of amastigotes.
Article
Biochemistry & Molecular Biology
Roberto Docampo, Anibal Eugenio Vercesi
Summary: This review discusses the mechanisms of mitochondrial oxidant generation and removal, and the involvement of Ca2+ in trypanosome cell death, highlighting the need for further studies on ROS generation, defense mechanisms, and mitochondrial permeability transition pore in trypanosomatids.
ANTIOXIDANTS & REDOX SIGNALING
(2022)
Article
Microbiology
Fatma Gomaa, Zhu-Hong Li, David J. Beaudoin, Heba Alzan, Peter R. Girguis, Roberto Docampo, Virginia P. Edgcomb
Summary: Developing transfection protocols for marine protists is an emerging field. In this study, we successfully developed a CRISPR/Cas9 editing protocol for Bodo saltans and identified two mechanisms for repairing double-strand breaks in this organism.
ENVIRONMENTAL MICROBIOLOGY
(2022)
Review
Microbiology
Roberto Docampo, Guozhong Huang
Summary: Acidocalcisomes are organelles found in bacteria and eukaryotes. They play important roles in autophagy, ion and phosphorus storage, osmoregulation, pH homeostasis, and pathogenesis. Recent studies have also revealed their involvement in phosphate utilization, calcium ion signaling, and bioenergetics.
JOURNAL OF EUKARYOTIC MICROBIOLOGY
(2022)
Article
Immunology
Thaise Lara Teixeira, Miguel Angel Chiurillo, Noelia Lander, Cassiano Costa Rodrigues, Thiago Souza Onofre, Eden Ramalho Ferreira, Camila Miyagui Yonamine, Julia de Gouveia Santos, Renato Arruda Mortara, Claudio Vieira da Silva, Jose Franco da Silveira
Summary: P21 is an immunomodulatory protein in Trypanosoma cruzi that plays crucial roles in host cell invasion, internalization, and amastigote proliferation.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2022)
Article
Chemistry, Medicinal
Alejandro Pezza, Luis E. Tavernelli, Victoria L. Alonso, Virginia Perdomo, Raquel Gabarro, Rab Prinjha, Elvio Rodriguez Araya, Inmaculada Rioja, Roberto Docampo, Felix Calderon, Julio Martin, Esteban Serra
Summary: TcBDF2 is an important bromodomain protein in T. cruzi, playing a crucial role in infection, replication, and differentiation of the parasite, as well as pathogenesis.
ACS INFECTIOUS DISEASES
(2022)
Article
Cell Biology
Mayara S. Bertolini, Roberto Docampo
Summary: In this study, we generated TcMICU1/TcMICU2 double knockout cell lines using CRISPR/Cas9 genome editing and found that these knockout cell lines were unable to uptake calcium at low extramitochondrial calcium concentrations. Further investigation revealed that Mg2+ negatively regulates the mitochondrial calcium uptake mediated by MCUC.
Editorial Material
Cell Biology
Roberto Docampo
Article
Microbiology
Nuria W. Negrao, Logan P. Crowe, Brian S. Mantilla, Rodrigo P. Baptista, Sharon King-Keller, Guozhong Huang, Roberto Docampo
Summary: Trypanosoma brucei is the causative agent of African trypanosomiasis, and there is a need for new drugs due to limited treatment options and evidence of resistance. A phosphoinositide phospholipase C (TbPI-PLC-like) with an X and PDZ domain was discovered, similar to TbPI-PLC1. Recombinant TbPI-PLC-like does not hydrolyze phosphatidylinositol 4,5-bisphosphate (PIP2) and does not modulate TbPI-PLC1 activity, but its knockdown affects the proliferation of trypanosomes.
Review
Microbiology
Roberto Docampo, Corrella S. Detweiler
Summary: Acidocalcisomes are organelles with acidity and high content of polyphosphate and calcium, and they have various functions and interactions with other organelles.
MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS
(2023)