Is it possible to reconstruct an accurate cell lineage using CRISPR recorders?

Area of Science:

• Developmental Biology
• Genetics
• Computational Biology

Background:

• Cell lineages are crucial for understanding cell fate determination during development.
• Reconstructing cell lineages is challenging due to the large number of cells in multicellular organisms.
• CRISPR-induced mutations offer a potential method for heritable lineage tracing.

Purpose of the Study:

• To evaluate the accuracy of CRISPR-based lineage reconstruction methods.
• To identify key biological and technical parameters affecting CRISPR lineage recorder performance.
• To provide insights for optimizing future CRISPR lineage tracing strategies.

Main Methods:

• Computer simulations incorporating empirical CRISPR mutation data from *Drosophila*.
• Analysis of parameters including cell division rates, mutation outcomes, target dropouts, and sequencing strategies.
• Performance estimation of existing CRISPR recorder designs.

Main Results:

• Variable cell division rates, skewed mutational outcomes, target dropouts, and sequencing strategies significantly impact lineage reconstruction accuracy.
• Published CRISPR recorder designs have limitations under various conditions.
• The study quantifies the influence of different parameters on lineage tracing fidelity.

Conclusions:

• CRISPR-based lineage tracing holds promise but requires careful consideration of multiple factors for accurate reconstruction.
• Simulation-based analysis is essential for predicting and improving the performance of lineage tracing tools.
• Future CRISPR lineage recorder implementations can be optimized based on identified parameters to enhance accuracy and reliability.