MsgRNA · Modification of single guide (sg) RNA in CRISPR-Cas9 gene editing Tool
„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“
- Период
- 2019-11-01 → 2021-10-31
- Финансиране от ЕС
- 212 934 €
- Участници
- 1
- Схема
- MSCA-IF-EF-ST
Линиите свързват координатора с партньорите.
Накратко на български
Химичните модификации на CRISPR-Cas9 системата се изследват, за да се направи редактирането на гените по-точно и контролирано. Това помага при разработването на терапии за рак, метаболитни и невродегенеративни заболявания.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Modification of single guide (sg) RNA in CRISPR-Cas9 gene editing Tool
DNA stores the genetic information of almost all known organisms. The ability to precisely target DNA and alter its function has been a long-term goal of scientists. Until recently, methods to edit the genome were complex and unintuitive, but the discovery of RNA-guided CRISPR-Cas systems has transformed the situation. However, hurdles such as non-specific nucleobase editing, special information of gene editing programs, and controlling the cleavage of the desired gene are in front of the research in this field. Chemical manipulation of the system could prove vital in addressing these issues. This work is to expand the function of the system in order to ask better questions and simplify its use. The project is important for society because CRISPR is now in a stage of the human clinical trial as CRISPR-Cas9 gene-editing techniques are used in a variety of complex diseases, including cancer, neurodegenerative disorders, and genetic and metabolic diseases. So, modulation of the CRISPR gene-editing technique is the main challenge for society. In the project, we had used a simple technique to control the gene-editing mechanism. Using a multi-disciplinary approach combining innovative state-of-the-art probes and techniques, we characterized in depth the molecular mechanisms of gene editing CRISPR-Cas9 techniques.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
CRISPR/Cas9 has been extensively studied for genome editing, but its therapeutic application has been hampered by off-target effects. Site-specific modification of single guide (sg) RNA in CRISPR system is a potential means to expand the utility of CRISPR-Cas genome editing. To address this, and reduce of synthetic burden of sgRNA we propose a ‘click’ ligation method to synthesize sgRNA where two chemically modified oligonucleotides are joined together using CuAAC chemistry. The resultant artificial linkage is biomimetic, so it should not affect Cas9 activity. This method offers significantly higher DNA-targeting specificity (i.e. less off-target effects) and most importantly provides a cost-effective means to access thousands of synthetic sgRNAs. Imaging in CRISPR-Cas9 system using fluorescence in situ hybridisation (FISH) probes have enabled significant advancements in understanding genomic structure and transcriptional control. We will explore novel approaches to image CRISPR-Cas9 system using fluorophore-quencher pairs. The activation of fluorescence can be performed upon nuclear localisation using RNase H, and upon sgRNA binding to the target DNA. These methods will provide highly fluorescent sgRNAs for live-cell imaging, potentially much brighter compared to other methods. To control CRISPR off-target effects we will also design a light induced DNA damage method where cyanovinylcarbazole nucleoside or psolaren will be incorporated in the DNA-targeting RNA. Therefore, modification of sgRNA for CRISPR system can be used to address the biological limitations of CRISPR and expand its functionality. This will open up many avenues for future development and lead to more application-focused studies on therapeutic gene editing.
Оригинален текст от CORDIS (на английски).
Участници
- THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD · OxfordКоординаторОбединеното кралство
Връзки
Данни: CORDIS, © Европейски съюз