# Equipment Supplement for "Synthetic genetic systems for rapid biomolecular evolution in vivo"

> **NIH NIH R35** · UNIVERSITY OF CALIFORNIA-IRVINE · 2021 · $71,950

## Abstract

I. Project Summary/Abstract
Over the past five years, my lab has made significant strides in the development of genetic systems capable of
driving the rapid mutation and evolution of user-selected genes of interest in vivo. These systems have allowed
us and others to quickly and scalably evolve enzymes, proteins, and antibodies to address a range of problems
spanning from studying drug resistance to creating affinity reagents on demand. These systems have also begun
to allow us to use rapid mutational accumulation as a method for tracing cell lineage in developing animals. One
of our key accomplishments has been the invention of an orthogonal DNA replication system (OrthoRep). In
OrthoRep, an error-prone orthogonal DNA polymerase (DNAP) exclusively replicates a special cytosolic plasmid
encoding only genes of interest (GOIs), driving their continuous evolution fully in vivo. The parent MIRA grant
will integrate our lab’s work on OrthoRep and support its further development and application in the next five
years. In particular, we will grow the core OrthoRep technology in order to accelerate GOI evolution in yeast
even more than we currently have, attempt to establish OrthoRep in mammalian cells in order to extend the
range of problems OrthoRep can directly address, apply OrthoRep to the engineering of bespoke Cas9s to
extend the range and efficacy of targeting, apply OrthoRep to improve the capabilities of a lineage tracing tool
developed by my lab, and apply OrthoRep to the generation of mutually orthogonal collections of aminoacyl-
tRNA synthetase (aaRS)/tRNA pairs to support genetic code expansion efforts also ongoing in my lab. We hope
the set of activities proposed for this MIRA will solidify OrthoRep as an exceptionally powerful genetic system for
evolving enzymes and proteins capable of solving high-reward problems in the chemical, biological, and
biomedical sciences.

## Key facts

- **NIH application ID:** 10387760
- **Project number:** 3R35GM136297-02S1
- **Recipient organization:** UNIVERSITY OF CALIFORNIA-IRVINE
- **Principal Investigator:** Chang C Liu
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $71,950
- **Award type:** 3
- **Project period:** 2020-06-01 → 2025-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10387760

## Citation

> US National Institutes of Health, RePORTER application 10387760, Equipment Supplement for "Synthetic genetic systems for rapid biomolecular evolution in vivo" (3R35GM136297-02S1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10387760. Licensed CC0.

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