# Development and Application of New Genome Editing Tools for the Functional Investigation of Genetic Variants of Uncertain Significance

> **NIH NIH R35** · UNIVERSITY OF CALIFORNIA, SAN DIEGO · 2023 · $356,242

## Abstract

Komor – Project Summary/Abstract – “Development and Application of New Genome Editing Tools for
the Functional Investigation of Genetic Variants of Uncertain Significance”
My research program aims to combat the variant interpretation problem that looms over the field of precision
medicine: out of 4.6 million missense variants identified in the Genome Aggregation Database, over half are
admitted to be variants of uncertain significance (VUS). New methods to enable the interpretation and functional
characterization of these VUS would not only enhance the efficacy of current therapies by better informing patient
selection strategies, but also accelerate the development of new approaches to combat diseases with a genetic
component. Targeted genome editing, the introduction of a specific modification in genomic DNA, has the
potential to allow researchers to study and better understand mechanisms of human genetic diseases, but
traditional genome editing methods (including CRISPR-Cas9) suffer from modest genome editing efficiencies as
well as unwanted gene alterations, particularly when attempting to introduce point mutations due to their reliance
on double-stranded DNA breaks (DSBs). Recently, I developed a class of genome editing agents called base
editors that does not involve DSBs, but rather uses a catalytically inactive Cas9 tethered to a single-stranded
DNA modifying enzyme to directly chemically modify target nucleobases in genomic DNA. Two classes of editors
currently exist, which use cytosine and adenine deamination chemistries to catalyze the conversion of C•G base
pairs to T•A (CBEs), and A•T base pairs to G•C (ABEs), respectively. My research program involves both the
development of new base editor methodologies, as well as the utilization of currently available base editor tools
to functionally interrogate VUS. Direction 1 research aims to develop new base editors capable of facilitating
new point mutations using computationally-aided directed evolution. The resulting tools will be of broad interest
to the scientific community as they will enable researchers to cleanly and efficiently install additional types of
point mutations into the genome of living cells, enabling the study and potential treatment of human genetic
diseases. Direction 2 research endeavors to initiate the first investigation into the pathogenicity of co-occurring
VUS (i.e. when a given individual has two or more VUS in their genome) through the development of orthogonal
base editing. Bioinformatic analyses of ours suggest that the clinical interpretation of missense variants is being
convoluted by their frequent co-occurrence with other uninterpreted variants, and the development of orthogonal
base editing will allow us to functionally interrogate these co-occurring variants and assess their contribution to
human genetic diseases. Finally, Direction 3 research proposes the development of high-throughput base
editing, which will allow for the functional investigation of ...

## Key facts

- **NIH application ID:** 10690519
- **Project number:** 5R35GM138317-04
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN DIEGO
- **Principal Investigator:** Alexis C. Komor
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $356,242
- **Award type:** 5
- **Project period:** 2020-09-01 → 2025-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10690519

## Citation

> US National Institutes of Health, RePORTER application 10690519, Development and Application of New Genome Editing Tools for the Functional Investigation of Genetic Variants of Uncertain Significance (5R35GM138317-04). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10690519. Licensed CC0.

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