ArticleG3 (Bethesda, Md.)2018
Double Selection Enhances the Efficiency of Target-AID and Cas9-Based Genome Editing in Yeast.
Article in G3 (Bethesda, Md.), 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- An Ammonium Transporter Gene Contributes to the Aggressiveness of the Dutch Elm Disease PathogenJournal of fungi (Basel, Switzerland) · 2026Article
- High-Efficiency Multiplexed Cytosine Base Editors for Natural Product Synthesis inACS synthetic biology · 2023Article
- Asymmetrical dose responses shape the evolutionary trade-off between antifungal resistance and nutrient use.Nature ecology & evolution · 2022Article
- Research progress of pathway and genome evolution in microbes.Synthetic and systems biotechnology · 2022Article
- High-Throughput Gene Mutagenesis Screening Using Base Editing.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Target-AID-Mediated Multiplex Base Editing in Porcine Fibroblasts.Animals : an open access journal from MDPI · 2021Article
- Universal toxin-based selection for precise genome engineering in human cells.Nature communications · 2021Article
- CRISPR-Cas12a system in fission yeast for multiplex genomic editing and CRISPR interference.Nucleic acids research · 2020Article
- Advances in Molecular Tools and In Vivo Models for the Study of Human Fungal Pathogenesis.Microorganisms · 2020Review
- CRISPR/Cas9 Gene Editing: An Unexplored Frontier for Forest Pathology.Frontiers in plant science · 2020Review
- Paralog dependency indirectly affects the robustness of human cells.Molecular systems biology · 2019Article
- Reshuffling yeast chromosomes with CRISPR/Cas9.PLoS genetics · 2019Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
CRISPR-Cas9 loss of function (LOF) and base editing screens are powerful tools in genetics and genomics. Yeast is one of the main models in these fields, but has only recently started to adopt this new toolkit for high throughput experiments. We developed a double selection strategy based on co-selection that increases LOF mutation rates using the Target-AID base editor. We constructed the pDYSCKO vector, which is amenable to high throughput double selection experiments, and show that the improvement in Target-AID efficiency generalizes across loci. Using modeling, we show that this improvement in efficiency provides the required increased in detection power to measure the fitness effects of thousands of mutations in typical yeast pooled screens. We show that double selection can also improve Cas9 mediated LOF rates, but that this multiplex genome editing causes programmable chromosomal translocations at high frequency. This suggests that multiplex LOF editing should be performed with caution and that base-editors could be preferable tools for some screens in yeast. Base editing using double selection is simple and straightforward and provides an alternative to homology directed repair based high throughput variant strain construction methods.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.