دورية أكاديمية
Comprehensive Screening of a Light-Inducible Split Cre Recombinase with Domain Insertion Profiling.
| العنوان: | Comprehensive Screening of a Light-Inducible Split Cre Recombinase with Domain Insertion Profiling. |
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| المؤلفون: | Tague N; Department of Biomedical Engineering, Boston University, Boston, Massachusetts 02215, United States.; Biological Design Center, Boston University, Boston, Massachusetts 02215, United States., Andreani V; Department of Biomedical Engineering, Boston University, Boston, Massachusetts 02215, United States.; Biological Design Center, Boston University, Boston, Massachusetts 02215, United States., Fan Y; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21218, United States., Timp W; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21218, United States.; Department of Molecular Biology and Genetics, Johns Hopkins University, Baltimore, Maryland 21218, United States., Dunlop MJ; Department of Biomedical Engineering, Boston University, Boston, Massachusetts 02215, United States.; Biological Design Center, Boston University, Boston, Massachusetts 02215, United States. |
| المصدر: | ACS synthetic biology [ACS Synth Biol] 2023 Oct 20; Vol. 12 (10), pp. 2834-2842. Date of Electronic Publication: 2023 Oct 03. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S. |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 101575075 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2161-5063 (Electronic) Linking ISSN: 21615063 NLM ISO Abbreviation: ACS Synth Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, D.C. : American Chemical Society, c2012- |
| مواضيع طبية MeSH: | Genetic Engineering*/methods , Integrases*/genetics , Integrases*/metabolism, Bayes Theorem ; Protein Engineering ; Proteins |
| مستخلص: | Splitting proteins with light- or chemically inducible dimers provides a mechanism for post-translational control of protein function. However, current methods for engineering stimulus-responsive split proteins often require significant protein engineering expertise and the laborious screening of individual constructs. To address this challenge, we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out by using sequencing. We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on the split sites throughout the protein. To improve the accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures. Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest. |
| التعليقات: | Update of: bioRxiv. 2023 May 26;:. (PMID: 37293111) |
| معلومات مُعتمدة: | R01 AI102922 United States AI NIAID NIH HHS |
| فهرسة مساهمة: | Keywords: Bayesian inference; Cre recombinase; domain insertion profiling; optogenetics; protein engineering; split proteins |
| المشرفين على المادة: | EC 2.7.7.- (Cre recombinase) EC 2.7.7.- (Integrases) 0 (Proteins) |
| تواريخ الأحداث: | Date Created: 20231003 Date Completed: 20231023 Latest Revision: 20231024 |
| رمز التحديث: | 20240829 |
| DOI: | 10.1021/acssynbio.3c00328 |
| PMID: | 37788288 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 2161-5063 |
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| DOI: | 10.1021/acssynbio.3c00328 |