Evaluation of different methodology for gene editing of Clock genes (StEID1 and StLNK2) in potato (Solanum tuberosum )
(2025) MOBN03 20242Degree Projects in Molecular Biology
- Abstract
- Solanum tuberosum (Potato) is a cool-season, short-day crop that tuberize effectively at temperatures between 15°C-19°C and at a day length of about 12 hours. These environmental requirements pose a moderate challenge to potato cultivar in different region globally. In Northern Europe, summers are characterized by the prolonged daylight and temperature that frequently exceed 19℃. Such conditions can delay tuber initiation and stolon formation ultimately reducing number of tubers and overall yield.
For farmers, potato cultivation offers a significant economic potential due to its short growth cycle and high market demand. With more than 700 million people globally lacking adequate access to enough food, improving the potato productivity... (More) - Solanum tuberosum (Potato) is a cool-season, short-day crop that tuberize effectively at temperatures between 15°C-19°C and at a day length of about 12 hours. These environmental requirements pose a moderate challenge to potato cultivar in different region globally. In Northern Europe, summers are characterized by the prolonged daylight and temperature that frequently exceed 19℃. Such conditions can delay tuber initiation and stolon formation ultimately reducing number of tubers and overall yield.
For farmers, potato cultivation offers a significant economic potential due to its short growth cycle and high market demand. With more than 700 million people globally lacking adequate access to enough food, improving the potato productivity and adaptability is crucial. Potato is a high yielding and nutrient rich crop, containing essential carbohydrates, amino acids, dietary fibre, vitamins, and important minerals. As a result, it is considered as a complete food, for supporting human food access and overall well-being. Optimizing potato cultivars is essential to achieve consistent high yields and meet increasing production and nutritional demands worldwide.
In this thesis, we focused on the genome editing of clock-associated genes involved in photoperiod perception, specifically StEID1 and StLNK2, in the potato cultivar ‘Desirée’. The primary objective was to reprogram the plant’s circadian clock to improve its adaptation to long-day conditions, which are critical for enhancing tuberization and maximizing yield. To achieve this, we employed advanced CRISPR-Cas technologies to introduce targeted mutations. Notably, our results demonstrate the successful application of homology-directed repair (HDR) to precisely modify these genes. Furthermore, we achieved efficient and simultaneous targeting of both genes, highlighting the potential of multiplex editing strategies for complex trait improvement in potato. (Less) - Popular Abstract
- CLOCK GENE EDITING – USING CRISPR Cas
Potato is a major global food crop, but its growth is sensitive to environmental factors like temperature and day length. Most of the essential activities in a plant, like growth, flowering, and seed or tuber production are controlled by its internal "clock," through the circadian rhythm. This clock helps plants keep track of time and adjust to changes in the environment, like longer days in summer or shorter days in winter. For a plant to form tubers it’s important that this internal clock stays in sync with the outside environment. Two important genes, called LNK2 and EID1, help the plant sense light and integrate to clock, that control the internal signals that decide when it’s time to start... (More) - CLOCK GENE EDITING – USING CRISPR Cas
Potato is a major global food crop, but its growth is sensitive to environmental factors like temperature and day length. Most of the essential activities in a plant, like growth, flowering, and seed or tuber production are controlled by its internal "clock," through the circadian rhythm. This clock helps plants keep track of time and adjust to changes in the environment, like longer days in summer or shorter days in winter. For a plant to form tubers it’s important that this internal clock stays in sync with the outside environment. Two important genes, called LNK2 and EID1, help the plant sense light and integrate to clock, that control the internal signals that decide when it’s time to start forming tubers. Shorter days help the plant to grow tubers, while longer days can slow down or stop this process.
The purpose of editing these genes is to study and manipulate traits that are critical for crop productivity. One such trait is photoperiod sensitivity, which means how a plant responds to day length, which strongly influences tuberization time and yield stability across different latitudes. Genes involved in photoperiod pathways, such as LNK2, EID1and andCDF1 in potato, act as key regulators that help the plant detect light and clock to decide when to initiate tuberization. Modifying these genes can help develop varieties better suited to specific environments and achieve better yield. This project used a tool called CRISPR-Cas, which works like a pair of genetic scissors. It can find a specific spot in the DNA with the help of a guide (RNA) and then make a precise cut at targeted location. After the cut, the cell tries to fix the damage. Sometimes, the repair isn’t perfect, which can cause a small change or "mutation" in the DNA. In other cases, template was provided a sequence of nucleotides the cell can copy, so the repair creates a specific, intended mutation based on that template. Guides designed for this project were attached to the gene scissor and then introduced into potato cells.
Guide was designed to cause changes in a specific spot of EID1 and LNK2 gene. The objective was then to compare the changes of each repair and see which one caused more changes and which one was better at turning off the genes. To do this, the DNA of the potato cells needs to be extracted and then sequenced. Different proteins like Cas-9 and Cas-12a, were tested to see which one does a better job at making accurate changes to the plant’s DNA. Their editing performance was carefully compared.
Sequences showed that the modification was achieved. Together with which it shows that two genes can be targeted at the same time. The changes seen was the same desired change which was introduced into the potato cells at smaller ratio.
Future work should aim to boost editing accuracy by improving guide design and introducing double cuts at gene loci, making results more improved with intended mutation and reliable for plant research and breeding.
Master’s degree project in Molecular biology/ 60 Credits. Department of biology, Lund university
Advisor: Per Hofvander (Department of Plant Breeding, SLU) (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9211911
- author
- Arani Suresh, Haripriyaa
- supervisor
- organization
- course
- MOBN03 20242
- year
- 2025
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 9211911
- date added to LUP
- 2025-09-08 14:18:33
- date last changed
- 2025-09-08 14:18:33
@misc{9211911,
abstract = {{Solanum tuberosum (Potato) is a cool-season, short-day crop that tuberize effectively at temperatures between 15°C-19°C and at a day length of about 12 hours. These environmental requirements pose a moderate challenge to potato cultivar in different region globally. In Northern Europe, summers are characterized by the prolonged daylight and temperature that frequently exceed 19℃. Such conditions can delay tuber initiation and stolon formation ultimately reducing number of tubers and overall yield.
For farmers, potato cultivation offers a significant economic potential due to its short growth cycle and high market demand. With more than 700 million people globally lacking adequate access to enough food, improving the potato productivity and adaptability is crucial. Potato is a high yielding and nutrient rich crop, containing essential carbohydrates, amino acids, dietary fibre, vitamins, and important minerals. As a result, it is considered as a complete food, for supporting human food access and overall well-being. Optimizing potato cultivars is essential to achieve consistent high yields and meet increasing production and nutritional demands worldwide.
In this thesis, we focused on the genome editing of clock-associated genes involved in photoperiod perception, specifically StEID1 and StLNK2, in the potato cultivar ‘Desirée’. The primary objective was to reprogram the plant’s circadian clock to improve its adaptation to long-day conditions, which are critical for enhancing tuberization and maximizing yield. To achieve this, we employed advanced CRISPR-Cas technologies to introduce targeted mutations. Notably, our results demonstrate the successful application of homology-directed repair (HDR) to precisely modify these genes. Furthermore, we achieved efficient and simultaneous targeting of both genes, highlighting the potential of multiplex editing strategies for complex trait improvement in potato.}},
author = {{Arani Suresh, Haripriyaa}},
language = {{eng}},
note = {{Student Paper}},
title = {{Evaluation of different methodology for gene editing of Clock genes (StEID1 and StLNK2) in potato (Solanum tuberosum )}},
year = {{2025}},
}