Codon-Mutant Libraries of Protein-Coding Genes

Technology Overview

Random mutant libraries of genes are used in a wide variety of molecular biology and bioengineering applications. Current techniques (such as error-prone PCR and its variants) are well suited for introducing mutations at the nucleotide level. PCR with oligonucleotides containing degenerate bases at certain positions are well suited for creating randomized codons at a small number of positions, particularly if these positions are nearly contiguous in primary sequence. However, there are no standard techniques for randomizing codons uniformly throughout a gene at a low rate of mutagenesis. Dr. Bloom has devised an approach to create random codon mutations distributed uniformly across a gene. The Bloom Lab has successfully applied this technique to randomly mutate all codon positions (except the start and stop codons) in the 1.5 kb influenza nucleoprotein gene as proof of concept.

Applications

•       Vaccine development
•       Directed evolution and protein manufacturing
•       Microbiome analysis for cancer therapies

Advantages

•       Expands the genetic space that can be searched in mutant libraries
•       Introduces mutations at a rate of 2-3 codon mutations per gene
•         Decreases rate of insertions and deletions introduced to less than 0.1 indels per gene

Patent Information

• 10,479,989

Market Overview

•  The vaccines market is estimated to be worth USD 77.6 billion in 2023 and to expand to USD $93.8 billion by 2028 with a CAGR of 3.9 %.

Investigator Overview

• Jesse Bloom, PhD - Basic Science and Public Health Divisions