Random Codon Mutant Libraries Created by PCR Using Synthetic Nucleotides

Description:

Codon-Mutant Libraries of Protein-Coding Genes

 

Technique that mutates entire genes at the codon level at a controlled rate, compared to existing error-prone PCR methods that achieve this on the nucleotide level

 

 

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.

 

 

<ul>

      <li>Vaccine development</li>

      <li>Directed evolution and protein manufacturing</li>

      <li>Microbiome analysis for cancer therapies</li>

</ul>

     

 

<ul>

      <li>Expands the genetic space that can be searched in mutant libraries</li>

      <li>Introduces mutations at a rate of 2-3 codon mutations per gene</li>

       <li>Decreases rate of insertions and deletions introduced to less than 0.1 indels per gene </li>

</ul>

 

 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 %.

 

<ul>

      <li>Jesse Bloom, PhD - Basic Science and Public Health Divisions</li>

</ul>

 

Preclinical <em>in vitro</em>

 

10,479,989

 

13-033 Bloom_ Codon-Mutant Libraries of Protein-Coding Genes.pdf

 

| influenza| mutant |codon | Library

 

Codon-Mutant-Libraries-Protein-Coding-Genes-13-033

 

 

 

 

Patent Information:
Category(s):
Therapeutic
For Information, Contact:
Patrick Shelby
Director, Technology Management
Fred Hutchinson Cancer Research Center
206-667-5821
jpshelby@fredhutch.org
Inventors:
Jesse Bloom
Keywords:
Assay / Method
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