PloS one, 2018-04, Vol.13 (4), p.e0195558-e0195558
2018
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Details
- Autor(en) / Beteiligte
- Titel
- Membrane permeabilizing amphiphilic peptide delivers recombinant transcription factor and CRISPR-Cas9/Cpf1 ribonucleoproteins in hard-to-modify cells
- Ist Teil von
- PloS one, 2018-04, Vol.13 (4), p.e0195558-e0195558
- Ort / Verlag
- United States: Public Library of Science
- Erscheinungsjahr
- 2018
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Delivery of recombinant proteins to therapeutic cells is limited by a lack of efficient methods. This hinders the use of transcription factors or Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) ribonucleoproteins to develop cell therapies. Here, we report a soluble peptide designed for the direct delivery of proteins to mammalian cells including human stem cells, hard-to-modify primary natural killer (NK) cells, and cancer cell models. This peptide is composed of a 6x histidine-rich domain fused to the endosomolytic peptide CM18 and the cell penetrating peptide PTD4. A less than two-minute co-incubation of 6His-CM18-PTD4 peptide with spCas9 and/or asCpf1 CRISPR ribonucleoproteins achieves robust gene editing. The same procedure, co-incubating with the transcription factor HoxB4, achieves transcriptional regulation. The broad applicability and flexibility of this DNA- and chemical-free method across different cell types, particularly hard-to-transfect cells, opens the way for a direct use of proteins for biomedical research and cell therapy manufacturing.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1932-6203eISSN: 1932-6203DOI: 10.1371/journal.pone.0195558Titel-ID: cdi_plos_journals_2021717287
- Format
- –
- Schlagworte
- Animals, Biology and Life Sciences, Cancer, Cell culture, Cells, Cultured, Cellular therapy, Cerebral Cortex - cytology, Cerebral Cortex - metabolism, Cricetulus, CRISPR, CRISPR-Cas Systems, Cytosol - metabolism, Deoxyribonucleic acid, DNA, Endocytosis, Engineering and Technology, Escherichia coli, Gene Editing - methods, Gene expression, Gene regulation, Genetic modification, Genome editing, Health aspects, Heparan sulfate, Histidine, HOXB4 protein, Humans, Incubation, Mammalian cells, Medical research, Medicine, Experimental, Membranes, Methods, Mice, Myoblasts - cytology, Myoblasts - metabolism, Natural killer cells, Peptides, Protein engineering, Proteins, Rats, Recombinant proteins, Recombinant Proteins - administration & dosage, Recombinant Proteins - metabolism, Research and analysis methods, Ribonucleoproteins, Stem cells, Stem Cells - cytology, Stem Cells - metabolism, Transcription factors, Transcription Factors - administration & dosage, Transcription Factors - metabolism