CORALINA: a universal method for the generation of gRNA libraries for CRISPR-based screening
| العنوان: | CORALINA: a universal method for the generation of gRNA libraries for CRISPR-based screening |
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| المؤلفون: | Christopher T. Breunig, Christiane Fuchs, Karolina Worf, Stephan Beck, Valentin Baumann, Maximilian F. Wiesbeck, Stefan H. Stricker, Lukas Hutter, Anna Köferle, Magdalena Götz, Javier Herrero |
| المصدر: | BMC Genomics Europe PubMed Central BMC Genomics 17:917 (2016) |
| بيانات النشر: | Springer Science and Business Media LLC, 2016. |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, Genomics, Computational biology, Biology, Mice, 03 medical and health sciences, Epigenome editing, Genetics, Animals, Humans, Epigenetic engineering, CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats, Genomic library, Genome-wide, Cas9, Elongated protospacer, Gene Library, Epigenomics, Methodology Article, Reproducibility of Results, genomic DNA, 030104 developmental biology, Genetic engineering, gRNA library, Elongated Protospacer, Epigenetic Engineering, Epigenome Editing, Genetic Engineering, Grna Library, CRISPR-Cas Systems, DNA microarray, RNA, Guide, Kinetoplastida, Biotechnology |
| الوصف: | Background The bacterial CRISPR system is fast becoming the most popular genetic and epigenetic engineering tool due to its universal applicability and adaptability. The desire to deploy CRISPR-based methods in a large variety of species and contexts has created an urgent need for the development of easy, time- and cost-effective methods enabling large-scale screening approaches. Results Here we describe CORALINA (comprehensive gRNA library generation through controlled nuclease activity), a method for the generation of comprehensive gRNA libraries for CRISPR-based screens. CORALINA gRNA libraries can be derived from any source of DNA without the need of complex oligonucleotide synthesis. We show the utility of CORALINA for human and mouse genomic DNA, its reproducibility in covering the most relevant genomic features including regulatory, coding and non-coding sequences and confirm the functionality of CORALINA generated gRNAs. Conclusions The simplicity and cost-effectiveness make CORALINA suitable for any experimental system. The unprecedented sequence complexities obtainable with CORALINA libraries are a necessary pre-requisite for less biased large scale genomic and epigenomic screens. Electronic supplementary material The online version of this article (doi:10.1186/s12864-016-3268-z) contains supplementary material, which is available to authorized users. |
| وصف الملف: | application/pdf |
| تدمد: | 1471-2164 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2d1186f4c15d75345a4152c56ffc0e2c https://doi.org/10.1186/s12864-016-3268-z |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....2d1186f4c15d75345a4152c56ffc0e2c |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 14712164 |
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