Developmental barcoding of whole mouse via homing CRISPR

Area of Science:

• Developmental Biology
• Genetics
• Molecular Biology

Background:

• In vivo barcoding is crucial for understanding biological information, including developmental lineages.
• Previous applications of nuclease-induced mutation barcoding in mammalian systems have been limited.
• CRISPR-based lineage tracing offers potential for high-resolution biological recording.

Purpose of the Study:

• To establish a versatile in vivo barcoding platform in mice.
• To enable high-throughput lineage tracing and developmental studies in mammals.
• To reconstruct early developmental lineages and investigate brain axis development.

Main Methods:

• Utilized multiple homing guide RNAs for CRISPR-mediated mutagenesis in vivo.
• Activated barcoding upon conception and continued mutagenesis throughout gestation.
• Generated exponentially diverse barcodes through combined mutations.
• Applied barcoding for post hoc reconstruction of early lineages and brain development analysis.

Main Results:

• Successfully created developmentally barcoded mice with lineage-specific mutations.
• Achieved reliable reconstruction of the earliest mammalian lineages.
• Enabled investigation into the developmental processes of brain axis formation.
• Demonstrated the platform's capability for high-resolution lineage tracing.

Conclusions:

• The developed in vivo barcoding system is an enabling and versatile platform for mammalian research.
• This technology significantly advances lineage tracing capabilities in mouse models.
• The system provides new avenues for studying developmental biology and disease mechanisms.