Related Experiment Video
Updated: Mar 31, 2026

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
Published on: December 11, 2020
Improving a Synechocystis-based photoautotrophic chassis through systematic genome mapping and validation of neutral
Filipe Pinto1, Catarina C Pacheco2, Paulo Oliveira2
1i3S-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal IBMC-Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto 4150-180, Portugal Faculdade de Ciências, Departamento de Biologia, Universidade do Porto, Porto 4150-171, Portugal.
Abstract:
The use of microorganisms as cell factories frequently requires extensive molecular manipulation. Therefore, the identification of genomic neutral sites for the stable integration of ectopic DNA is required to ensure a successful outcome. Here we describe the genome mapping and validation of five neutral sites in the chromosome of Synechocystis sp. PCC 6803, foreseeing the use of this cyanobacterium as a photoautotrophic chassis. To evaluate the neutrality of these loci, insertion/deletion mutants were produced, and to assess their functionality, a synthetic green fluorescent reporter module was introduced. The constructed integrative vectors include a BioBrick-compatible multiple cloning site insulated by transcription terminators, constituting robust cloning interfaces for synthetic biology approaches. Moreover, Synechocystis mutants (chassis) ready to receive purpose-built synthetic modules/circuits are also available. This work presents a systematic approach to map and validate chromosomal neutral sites in cyanobacteria, and that can be extended to other organisms.
Related Concept Videos
CRISPR/Cas9 Genome Editing
Conservative Site-specific Recombination and Phase Variation
The recognition sites for Cre recombinase called LoxP...
CRISPR
CRISPR and crRNAs
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...

