Prime Editing Driven Functional Genomics: Bridging Genotype to Phenotype in the Post-Genomic Era.

Area of Science:

• Genomics
• Molecular Biology
• Genetic Engineering

Background:

• The post-genomic era has generated vast human genetic variation data, but most variants remain uncharacterized, hindering clinical applications.
• Current functional genomics methods like CRISPR-Cas9 screens cannot assess single nucleotide variants (SNVs) effectively.
• Bridging the genotype-to-phenotype gap requires high-throughput, precise functional genomics tools.

Purpose of the Study:

• To review the evolution of functional genomics screens from gene-level knockouts to advanced variant characterization.
• To highlight prime editing as a transformative platform for systematic variant functional analysis.
• To detail prime editing systems, their mechanisms, optimization, and delivery methods.

Main Methods:

• Review of CRISPR-Cas9 based methods including knockout, CRISPR interference/activation (CRISPRi/a), and multiplexed assays of variant effect (MAVEs).
• Discussion of base editors and prime editing for targeted genetic modifications.
• Analysis of saturation genome editing (SGE) approaches.

Main Results:

• Prime editing enables the systematic functional characterization of thousands of genetic variants with high precision.
• This technology overcomes limitations of previous methods in assessing SNVs and small indels.
• Prime editing systems have undergone progressive optimization in architecture, mechanism, and delivery.

Conclusions:

• Prime editing represents a new paradigm in functional genomics, enabling precise variant characterization at scale.
• This technology is poised to accelerate the diagnosis and treatment of genetic diseases.
• The systematic functional analysis of variants using prime editing will advance precision medicine.