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Joint genotypic and phenotypic outcome modeling improves base editing variant effect quantification
CRISPR base editing screens are powerful tools for studying disease-associated variants at scale. However, the efficiency and precision of base editing perturbations vary, confounding the assessment of variant-induced phenotypic effects. Here, we provide an integrated pipeline that improves the esti...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10508837/ https://www.ncbi.nlm.nih.gov/pubmed/37732177 http://dx.doi.org/10.1101/2023.09.08.23295253 |
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author | Ryu, Jayoung Barkal, Sam Yu, Tian Jankowiak, Martin Zhou, Yunzhuo Francoeur, Matthew Phan, Quang Vinh Li, Zhijian Tognon, Manuel Brown, Lara Love, Michael I. Lettre, Guillaume Ascher, David B. Cassa, Christopher A. Sherwood, Richard I. Pinello, Luca |
author_facet | Ryu, Jayoung Barkal, Sam Yu, Tian Jankowiak, Martin Zhou, Yunzhuo Francoeur, Matthew Phan, Quang Vinh Li, Zhijian Tognon, Manuel Brown, Lara Love, Michael I. Lettre, Guillaume Ascher, David B. Cassa, Christopher A. Sherwood, Richard I. Pinello, Luca |
author_sort | Ryu, Jayoung |
collection | PubMed |
description | CRISPR base editing screens are powerful tools for studying disease-associated variants at scale. However, the efficiency and precision of base editing perturbations vary, confounding the assessment of variant-induced phenotypic effects. Here, we provide an integrated pipeline that improves the estimation of variant impact in base editing screens. We perform high-throughput ABE8e-SpRY base editing screens with an integrated reporter construct to measure the editing efficiency and outcomes of each gRNA alongside their phenotypic consequences. We introduce BEAN, a Bayesian network that accounts for per-guide editing outcomes and target site chromatin accessibility to estimate variant impacts. We show this pipeline attains superior performance compared to existing tools in variant classification and effect size quantification. We use BEAN to pinpoint common variants that alter LDL uptake, implicating novel genes. Additionally, through saturation base editing of LDLR, we enable accurate quantitative prediction of the effects of missense variants on LDL-C levels, which aligns with measurements in UK Biobank individuals, and identify structural mechanisms underlying variant pathogenicity. This work provides a widely applicable approach to improve the power of base editor screens for disease-associated variant characterization. |
format | Online Article Text |
id | pubmed-10508837 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-105088372023-09-20 Joint genotypic and phenotypic outcome modeling improves base editing variant effect quantification Ryu, Jayoung Barkal, Sam Yu, Tian Jankowiak, Martin Zhou, Yunzhuo Francoeur, Matthew Phan, Quang Vinh Li, Zhijian Tognon, Manuel Brown, Lara Love, Michael I. Lettre, Guillaume Ascher, David B. Cassa, Christopher A. Sherwood, Richard I. Pinello, Luca medRxiv Article CRISPR base editing screens are powerful tools for studying disease-associated variants at scale. However, the efficiency and precision of base editing perturbations vary, confounding the assessment of variant-induced phenotypic effects. Here, we provide an integrated pipeline that improves the estimation of variant impact in base editing screens. We perform high-throughput ABE8e-SpRY base editing screens with an integrated reporter construct to measure the editing efficiency and outcomes of each gRNA alongside their phenotypic consequences. We introduce BEAN, a Bayesian network that accounts for per-guide editing outcomes and target site chromatin accessibility to estimate variant impacts. We show this pipeline attains superior performance compared to existing tools in variant classification and effect size quantification. We use BEAN to pinpoint common variants that alter LDL uptake, implicating novel genes. Additionally, through saturation base editing of LDLR, we enable accurate quantitative prediction of the effects of missense variants on LDL-C levels, which aligns with measurements in UK Biobank individuals, and identify structural mechanisms underlying variant pathogenicity. This work provides a widely applicable approach to improve the power of base editor screens for disease-associated variant characterization. Cold Spring Harbor Laboratory 2023-09-10 /pmc/articles/PMC10508837/ /pubmed/37732177 http://dx.doi.org/10.1101/2023.09.08.23295253 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Ryu, Jayoung Barkal, Sam Yu, Tian Jankowiak, Martin Zhou, Yunzhuo Francoeur, Matthew Phan, Quang Vinh Li, Zhijian Tognon, Manuel Brown, Lara Love, Michael I. Lettre, Guillaume Ascher, David B. Cassa, Christopher A. Sherwood, Richard I. Pinello, Luca Joint genotypic and phenotypic outcome modeling improves base editing variant effect quantification |
title | Joint genotypic and phenotypic outcome modeling improves base editing variant effect quantification |
title_full | Joint genotypic and phenotypic outcome modeling improves base editing variant effect quantification |
title_fullStr | Joint genotypic and phenotypic outcome modeling improves base editing variant effect quantification |
title_full_unstemmed | Joint genotypic and phenotypic outcome modeling improves base editing variant effect quantification |
title_short | Joint genotypic and phenotypic outcome modeling improves base editing variant effect quantification |
title_sort | joint genotypic and phenotypic outcome modeling improves base editing variant effect quantification |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10508837/ https://www.ncbi.nlm.nih.gov/pubmed/37732177 http://dx.doi.org/10.1101/2023.09.08.23295253 |
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