dc.contributor.author | Cetin, Ronay | |
dc.contributor.author | Wegner, Martin | |
dc.contributor.author | Luwisch, Leah | |
dc.contributor.author | Saud, Sarada | |
dc.contributor.author | Achmedov, Tatjana | |
dc.contributor.author | Süsser, Sebastian | |
dc.contributor.author | Vera-Guapi, Antonella | |
dc.contributor.author | Müller, Konstantin | |
dc.contributor.author | Matthess, Yves | |
dc.contributor.author | Quandt, Eva | |
dc.contributor.author | Schaubeck, Simone | |
dc.contributor.author | Beisel, Chase L. | |
dc.contributor.author | Kaulich, Manuel | |
dc.date.accessioned | 2023-06-06T09:53:50Z | |
dc.date.available | 2023-06-06T09:53:50Z | |
dc.date.issued | 2023 | |
dc.identifier.citation | Cetin, Ronay; Wegner, Martin; Luwisch, Leah [et al.]. Optimized metrics for orthogonal combinatorial CRISPR screens. Scientific Reports, 2023, 13, 7405. Disponible en: <https://www.nature.com/articles/s41598-023-34597-8>. Fecha de acceso: 6 jun. 2023. DOI: 10.1038/s41598-023-34597-8 | ca |
dc.identifier.issn | 2045-2322 | ca |
dc.identifier.uri | http://hdl.handle.net/20.500.12328/3713 | |
dc.description.abstract | CRISPR-based gene perturbation enables unbiased investigations of single and combinatorial genotype-to-phenotype associations. In light of efforts to map combinatorial gene dependencies at scale, choosing an efficient and robust CRISPR-associated (Cas) nuclease is of utmost importance. Even though SpCas9 and AsCas12a are widely used for single, combinatorial, and orthogonal screenings, side-by-side comparisons remain sparse. Here, we systematically compared combinatorial SpCas9, AsCas12a, and CHyMErA in hTERT-immortalized retinal pigment epithelial cells and extracted performance-critical parameters for combinatorial and orthogonal CRISPR screens. Our analyses identified SpCas9 to be superior to enhanced and optimized AsCas12a, with CHyMErA being largely inactive in the tested conditions. Since AsCas12a contains RNA processing activity, we used arrayed dual-gRNAs to improve AsCas12a and CHyMErA applications. While this negatively influenced the effect size range of combinatorial AsCas12a applications, it enhanced the performance of CHyMErA. This improved performance, however, was limited to AsCas12a dual-gRNAs, as SpCas9 gRNAs remained largely inactive. To avoid the use of hybrid gRNAs for orthogonal applications, we engineered the multiplex SpCas9-enAsCas12a approach (multiSPAS) that avoids RNA processing for efficient orthogonal gene editing. | en |
dc.format.extent | 16 | ca |
dc.language.iso | eng | ca |
dc.publisher | Springer Nature | ca |
dc.relation.ispartof | Scientific Reports | ca |
dc.relation.ispartofseries | 13 | |
dc.relation.uri | https://www.nature.com/articles/s41598-023-34597-8 | ca |
dc.rights | This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. | en |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.subject.other | Tècniques biològiques | ca |
dc.subject.other | Enginyeria genètica | ca |
dc.subject.other | Projecció d'alt rendiment | ca |
dc.subject.other | Técnicas biológicas | es |
dc.subject.other | Ingeniería genética | es |
dc.subject.other | Proyección de alto impacto | es |
dc.subject.other | Biological techniques | en |
dc.subject.other | Genetic engineering | en |
dc.subject.other | High-throughput screening | en |
dc.title | Optimized metrics for orthogonal combinatorial CRISPR screens | en |
dc.type | info:eu-repo/semantics/article | ca |
dc.description.version | info:eu-repo/semantics/publishedVersion | ca |
dc.rights.accessLevel | info:eu-repo/semantics/openAccess | |
dc.embargo.terms | cap | ca |
dc.subject.udc | 61 | ca |
dc.identifier.doi | https://dx.doi.org/10.1038/s41598-023-34597-8 | ca |