دورية أكاديمية
Simple, fast and efficient iTOP-mediated delivery of CRISPR/Cas9 RNP in difficult-to-transduce human cells including primary T cells.
| العنوان: | Simple, fast and efficient iTOP-mediated delivery of CRISPR/Cas9 RNP in difficult-to-transduce human cells including primary T cells. |
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| المؤلفون: | Kholosy WM; Princess Máxima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, the Netherlands., Visscher M; NTrans Technologies BV, Heidelberglaan 7, 3584 CS Utrecht, the Netherlands., Ogink K; NTrans Technologies BV, Heidelberglaan 7, 3584 CS Utrecht, the Netherlands., Buttstedt H; NTrans Technologies BV, Heidelberglaan 7, 3584 CS Utrecht, the Netherlands., Griffin K; NTrans Technologies BV, Heidelberglaan 7, 3584 CS Utrecht, the Netherlands., Beier A; KNAW-Hubrecht Institute, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands., Gerlach JP; NTrans Technologies BV, Heidelberglaan 7, 3584 CS Utrecht, the Netherlands., Molenaar JJ; Princess Máxima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, the Netherlands., Geijsen N; KNAW-Hubrecht Institute, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands., de Boer M; NTrans Technologies BV, Heidelberglaan 7, 3584 CS Utrecht, the Netherlands., Chatsisvili A; NTrans Technologies BV, Heidelberglaan 7, 3584 CS Utrecht, the Netherlands. Electronic address: a.chatsisvili@uu.nl. |
| المصدر: | Journal of biotechnology [J Biotechnol] 2021 Sep 10; Vol. 338, pp. 71-80. Date of Electronic Publication: 2021 Jul 13. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Science Publishers Country of Publication: Netherlands NLM ID: 8411927 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4863 (Electronic) Linking ISSN: 01681656 NLM ISO Abbreviation: J Biotechnol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Amsterdam : Elsevier Science Publishers, c1984- |
| مواضيع طبية MeSH: | CRISPR-Cas Systems*/genetics , T-Lymphocytes*, Gene Editing ; Genome, Human ; HEK293 Cells ; Humans |
| مستخلص: | The advent of the CRISPR/Cas9 system has transformed the field of human genome engineering and has created new perspectives in the development of innovative cell therapies. However, the absence of a simple, fast and efficient delivery method of CRISPR/Cas9 into primary human cells has been limiting the progress of CRISPR/Cas9-based therapies. Here, we describe an optimized protocol for iTOP-mediated delivery of CRISPR/Cas9 in various human cells, including primary T cells, induced pluripotent stem cells (hiPSCs), Jurkat, ARPE-19 and HEK293 cells. We compare iTOP to other CRISPR/Cas9 delivery methods, such as electroporation and lipofection, and evaluate the corresponding gene-editing efficiencies and post-treatment cell viabilities. We demonstrate that the gene editing achieved by iTOP-mediated delivery of CRISPR/Cas9 is 40-95 % depending on the cell type, while post-iTOP cell viability remains high in the range of 70-95 %. Collectively, we present an optimized workflow for a simple, high-throughput and effective iTOP-mediated delivery of CRISPR/Cas9 to engineer difficult-to-transduce human cells. We believe that the iTOP technology® could contribute to the development of novel CRISPR/Cas9-based cell therapies. (Copyright © 2021 Elsevier B.V. All rights reserved.) |
| فهرسة مساهمة: | Keywords: CRISPR/Cas9; Gene editing; Primary human T cells; hiPSCs; iTOP |
| تواريخ الأحداث: | Date Created: 20210716 Date Completed: 20210924 Latest Revision: 20210924 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1016/j.jbiotec.2021.07.006 |
| PMID: | 34271056 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1873-4863 |
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| DOI: | 10.1016/j.jbiotec.2021.07.006 |