Research

Objectives

The Lig-Eng project, ‘Engineering DNA Ligases for Next-Generation-Sequencing’ aims to produce superior DNA ligase enzymes by discovering new highly-active variants from nature’s diversity and by engineering new features into naturally occurring ligase enzymes

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Work packages

WP1: Discovery, engineering and molecular biology

A bioinformatics approach is used to select novel DNA ligase genes from various microbes in different environments and discover improved activities. In silico design is used to make structure-guided changes to wild-type ligases to engineer improved properties.

The novel ligase genes are cloned into the expression host and screened for protein production under various expression conditions. Standard methods of protein expression and purification are used to produce large amounts of ligase enzymes for further characterization.

WP2: Activity testing

The ligase enzymes will be tested for ligase activity with different types of double- and single-stranded breaks with different reaction conditions to identify the best catalytic conditions and determine any other unusual or useful properties.

WP3: Structural characterization

The best-performing variants will be crystalized with the aim of obtaining high-resolution structures that could be used to rationalize activity and inform further enzyme-modification endeavors.

WP4: Application testing

The best-performing variants will be tested in real world applications

WP5: IP and commercialization

Patent application will be filed and commercialization opportunities will be explored with the goal to integrate our novel enzymes into a commercial library preparation kit.

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