CrisprBuildr: an open-source application for CRISPR-mediated genome engineering in Drosophila melanogaster

Area of Science:

• Molecular Biology
• Genomics
• Biotechnology

Background:

• CRISPR/Cas9 technology enables targeted genome editing for gene modification and characterization.
• Designing and cloning CRISPR constructs is often complex, time-consuming, and labor-intensive.
• Efficient tools are needed to facilitate CRISPR-based genome engineering workflows.

Purpose of the Study:

• To develop an open-source, web-based application, CrisprBuildr, to simplify the design of CRISPR/Cas9 genome engineering constructs.
• To assist researchers in designing guide RNAs and repair template vectors for gene editing.
• To create novel tagging vectors for optogenetic applications.

Main Methods:

• Development of CrisprBuildr, a web application for designing CRISPR/Cas9 guide RNAs and repair templates.
• Integration of the Drosophila melanogaster genome for targeted gene modification.
• Creation of new EGFP and mCherry tagging vectors incorporating the SspB-Q73R peptide for iLID-based optogenetics.

Main Results:

• CrisprBuildr successfully guides users through the design process, generating cloning maps for gene modifications.
• The application facilitates the creation of customized constructs for detailed gene and protein characterization.
• New tagging vectors suitable for optogenetic experiments were successfully generated.

Conclusions:

• CrisprBuildr significantly reduces the time and effort required for designing CRISPR/Cas9 genome engineering experiments.
• The developed tool supports efficient gene modification and protein characterization workflows.
• The application serves as a valuable resource for researchers utilizing CRISPR technology, with potential for adaptation to other genomes.