Gene editing laboratory (LEG)

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About the facility

The Gene Editing Laboratory (LEG) provides technical and scientific support to produce mammalian knockout cells using CRISPR/Cas9 technology. It provides services related to producing gene editing mechanisms and strategic planning experiments, with a focus on offering specialized training and support during the critical stages of the gene editing process.

The LEG facility is intended to expand Brazil’s capacity in gene editing, promoting technological autonomy and accelerating the generation of models for biomedical, pharmacological and biotechnological research.

How to submit research proposals

To use LEG you must submit a proposal through SAU Online. Calls will be announced every six months. If you have any questions, please contact CNPEM’s User Office at edu@cnpem.br or the team responsible for the facility at leg@lnbio.cnpem.br.

Equipment

  • Refrigerated microcentrifuge (Eppendorf);
  • Thermal cycler (BioRad);
  • ThermoMixer (Eppendorf);
  • NanodropOne UV VIS spectrophotometer (Thermo Scientific);
  • Horizontal electrophoresis system (Major Science);
  • Vertical electrophoresis system (BioRad);
  • Nucleofector 4D (Lonza);
  • Digital PCR QX600 (BioRad).

Main competencies and techniques

  • In silico analysis of target genes in mammalian genome banks;
  • Planning gene knockout strategies;
  • Guide cloning in plasmid vectors;
  • Design and in vitro transcription of guide RNA (sgRNA);
  • In vitro transcription of Cas9 mRNA;
  • Cloning-free production of sgRNA;
  • Preparing ribonucleoprotein complexes;
  • Quality control for reagents (concentration, integrity and functionality);
  • T7 Endonuclease I enzyme assay;
  • Technical support for optimizing transfection protocols (e.g. lipofection, electroporation, nucleofection);
  • Guidelines on selection and expansion of edited clones;
  • Support in standardizing validations, such as PCR, qPCR, Western blot, T7EI;
  • Discussing preliminary data and remote troubleshooting.

meet our team

Ângela Saito

Coordinator

Gene editing laboratory (LEG)
leg@lnbio.cnpem.br

Angelica Jardim Costa

Researcher

Gene editing laboratory (LEG)
leg@lnbio.cnpem.br

Gabriel Viliod Vieira, Karina Yumi Degaki, Raphael Morales Neto

Operations and technical support for users

Gene editing laboratory (LEG)
leg@lnbio.cnpem.br

General information

Contact LEG

leg@lnbio.cnpem.br