# A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC

## TL;DR

This paper introduces a new mutagenesis assay using specific DNA sequences to test how well a gene can tolerate mutations, which is important for biotechnology and drug development.

## Contribution

The novel mutagenesis assay uses NNT and NNC oligonucleotides to safely test gene mutability without causing harmful mutations.

## Key findings

- The NNT and NNC oligonucleotides prevent nonsense mutations and allow flexible insertion sizes.
- The ZeoR gene was confirmed to be highly tolerant to mutagenesis using this new assay.

## Abstract

Not all proteins are tolerable to mutations. Whether a specific protein can be a mutable target is of importance in the biotechnology and pharmaceutical industry. This study reported a novel mutagenesis assay using tandem NNT and NNC oligonucleotides to test the mutability of a candidate gene. These two tandem oligonucleotides avoid the risk of forming nonsense mutations and render flexibility of truncating or expanding the insertion size. As a reporter gene, ZeoR (zeocin resistance gene) was confirmed to have a high tolerance for mutagenesis by this new assay.