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Creating a modularly interactive CRISPR system

Vogel, Paul (2022) MOBN03 20212
Degree Projects in Molecular Biology
Popular Abstract
A modularly interactive CRISPR system

Climate change and the growing human population already pose severe challenges for food security and agricultural businesses around the globe. One key component on a way to a more sustainable agriculture that has the ability to adapt to changing environmental conditions could be the newly arisen genome editing. A form of ge- netic engineering that allows for precise edits without introducing foreign DNAs into the target organism. The core of genome editing is the gene scissor CRISPR/Cas9 that can be programmed to edit at a specific position in the genome. Here, we present the basis for a modular CRISPR system that has the potential to be more cost-efficient while providing a higher flexibility.

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A modularly interactive CRISPR system

Climate change and the growing human population already pose severe challenges for food security and agricultural businesses around the globe. One key component on a way to a more sustainable agriculture that has the ability to adapt to changing environmental conditions could be the newly arisen genome editing. A form of ge- netic engineering that allows for precise edits without introducing foreign DNAs into the target organism. The core of genome editing is the gene scissor CRISPR/Cas9 that can be programmed to edit at a specific position in the genome. Here, we present the basis for a modular CRISPR system that has the potential to be more cost-efficient while providing a higher flexibility.

DNA is a molecule that stores genetic information and functions as a blueprint for any living organism on earth. Manipulation of the genetic information can alter the organism. This has been indirectly done by humans to breed crop species with beneficial properties such as higher yield or better robustness. Plants that showed a preferred characteristic were selected for breeding. In this way plants with superior traits were created. Today the knowledge about genetics is being employed to directly facilitate this process. A change in the DNA, known to correlate with a beneficial trait, can be introduced into the plants genome by means of a range of breeding techniques.

Genome editing uses special proteins that can be programmed to target a specific position in the genome. The gene scissor CRISPR/Cas9 is the core of many genome editing technqiues, and allows us to cut at a defined position. Fusing other enzymes to Cas9 expands the functionality and enables us to insert or replace information. Delivering Cas9 as a protein directly into a plant cell, has the advantage that no foreign DNA is introduced, which lowers the risk for unforeseen side-effects. The production of the large Cas9 fusion proteins can be challenging and is currently a limiting factor.

In this thesis, we aimed to create a modular CRISPR system that separates the large fusion proteins into components. By adding small adapter proteins to the components we sought to build a plug-and-play system that provides both more cost-efficient protein production and higher flexibility. We were able to successfully produce the first components in bacteria and purify them for delivery to plant cells. Cas9 retained its cutting activity even when the adapter protein was added to it. Initial experiments showed that the components seemed to interact as expected. We hope that this technology expands the molecular toolbox and helps to engineer a new generation of crops.

Master’s Degree Project in Molecular Biology 60 credits 2022
Department of Biology, Lund University
Adviser: Per Hofvander
Sveriges lantbruketuniversitet (SLU) Alnarp (Less)
Please use this url to cite or link to this publication:
author
Vogel, Paul
supervisor
organization
course
MOBN03 20212
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
9103385
date added to LUP
2022-11-21 13:37:39
date last changed
2022-11-21 13:37:39
@misc{9103385,
  author       = {{Vogel, Paul}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Creating a modularly interactive CRISPR system}},
  year         = {{2022}},
}