دورية أكاديمية
CRISPR/Cas9 ribonucleoprotein-mediated knockin generation in hTERT-RPE1 cells.
| العنوان: | CRISPR/Cas9 ribonucleoprotein-mediated knockin generation in hTERT-RPE1 cells. |
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| المؤلفون: | Ghetti S; Armenise-Harvard Laboratory of Cell Division, Department of Cellular, Computational and Integrative Biology - CIBIO, University of Trento, Trento 38123, Italy., Burigotto M; Armenise-Harvard Laboratory of Cell Division, Department of Cellular, Computational and Integrative Biology - CIBIO, University of Trento, Trento 38123, Italy., Mattivi A; Armenise-Harvard Laboratory of Cell Division, Department of Cellular, Computational and Integrative Biology - CIBIO, University of Trento, Trento 38123, Italy., Magnani G; Armenise-Harvard Laboratory of Cell Division, Department of Cellular, Computational and Integrative Biology - CIBIO, University of Trento, Trento 38123, Italy., Casini A; Alia Therapeutics, Trento 38123, Italy., Bianchi A; Laboratory of Molecular Virology, Department of Cellular, Computational and Integrative Biology - CIBIO, University of Trento, Trento 38123, Italy., Cereseto A; Laboratory of Molecular Virology, Department of Cellular, Computational and Integrative Biology - CIBIO, University of Trento, Trento 38123, Italy., Fava LL; Armenise-Harvard Laboratory of Cell Division, Department of Cellular, Computational and Integrative Biology - CIBIO, University of Trento, Trento 38123, Italy. |
| المصدر: | STAR protocols [STAR Protoc] 2021 Mar 24; Vol. 2 (2), pp. 100407. Date of Electronic Publication: 2021 Mar 24 (Print Publication: 2021). |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Cell Press Country of Publication: United States NLM ID: 101769501 Publication Model: eCollection Cited Medium: Internet ISSN: 2666-1667 (Electronic) Linking ISSN: 26661667 NLM ISO Abbreviation: STAR Protoc |
| أسماء مطبوعة: | Original Publication: [Cambridge, MA] : Cell Press, [2020]- |
| مواضيع طبية MeSH: | CRISPR-Cas Systems/*genetics , Gene Knock-In Techniques/*methods , Retinal Pigment Epithelium/*cytology , Ribonucleoproteins/*genetics, Cell Line ; Gene Editing ; Humans |
| مستخلص: | hTERT-RPE1 cells are genetically stable near diploid cells widely used to model cell division, DNA repair, or ciliogenesis in a non-transformed context. However, poor transfectability and limited homology-directed repair capacity hamper their amenability to gene editing. Here, we describe a protocol for rapid and efficient generation of diverse homozygous knockins. In contrast to other approaches, this strategy bypasses the need for molecular cloning. Our approach can also be applied to a variety of cell types including cancer and induced pluripotent stem cells (iPSCs). Competing Interests: A. Cereseto is a founder and part of the Board of Directors of Alia Therapeutics Srl. A. Casini is a founder and an employee of Alia Therapeutics Srl. (© 2021 The Authors.) |
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| فهرسة مساهمة: | Keywords: CRISPR; Cell Biology |
| المشرفين على المادة: | 0 (Ribonucleoproteins) |
| تواريخ الأحداث: | Date Created: 20210415 Date Completed: 20220321 Latest Revision: 20240331 |
| رمز التحديث: | 20240331 |
| مُعرف محوري في PubMed: | PMC8025146 |
| DOI: | 10.1016/j.xpro.2021.100407 |
| PMID: | 33855309 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2666-1667 |
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| DOI: | 10.1016/j.xpro.2021.100407 |