Cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA for genome editing
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Suresh, B. | - |
dc.contributor.author | Ramakrishna, S. | - |
dc.contributor.author | Kim, H. | - |
dc.date.accessioned | 2022-07-14T23:40:47Z | - |
dc.date.available | 2022-07-14T23:40:47Z | - |
dc.date.created | 2021-05-11 | - |
dc.date.issued | 2017 | - |
dc.identifier.issn | 1064-3745 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/153275 | - |
dc.description.abstract | The clustered, regularly interspaced, short palindromic repeat (CRISPR)-associated (Cas) system represents an efficient tool for genome editing. It consists of two components: the Cas9 protein and a guide RNA. To date, delivery of these two components has been achieved using either plasmid or viral vectors or direct delivery of protein and RNA. Plasmid- and virus-free direct delivery of Cas9 protein and guide RNA has several advantages over the conventional plasmid-mediated approach. Direct delivery results in shorter exposure time at the cellular level, which in turn leads to lower toxicity and fewer off-target mutations with reduced host immune responses, whereas plasmid- or viral vector-mediated delivery can result in uncontrolled integration of the vector sequence into the host genome and unwanted immune responses. Cellpenetrating peptide (CPP), a peptide that has an intrinsic ability to translocate across cell membranes, has been adopted as a means of achieving efficient Cas9 protein and guide RNA delivery. We developed a method for treating human cell lines with CPP-conjugated recombinant Cas9 protein and CPP-complexed guide RNAs that leads to endogenous gene disruption. Here we describe a protocol for preparing an efficient CPP-conjugated recombinant Cas9 protein and CPP-complexed guide RNAs, as well as treatment methods to achieve safe genome editing in human cell lines. ? Springer Science+Business Media New York 2017. | - |
dc.language | 영어 | - |
dc.language.iso | en | - |
dc.publisher | Humana Press Inc. | - |
dc.title | Cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA for genome editing | - |
dc.type | Article | - |
dc.contributor.affiliatedAuthor | Ramakrishna, S. | - |
dc.identifier.doi | 10.1007/978-1-4939-6518-2_7 | - |
dc.identifier.scopusid | 2-s2.0-84994853550 | - |
dc.identifier.bibliographicCitation | Methods in Molecular Biology, v.1507, pp.81 - 94 | - |
dc.relation.isPartOf | Methods in Molecular Biology | - |
dc.citation.title | Methods in Molecular Biology | - |
dc.citation.volume | 1507 | - |
dc.citation.startPage | 81 | - |
dc.citation.endPage | 94 | - |
dc.type.rims | ART | - |
dc.type.docType | Book Chapter | - |
dc.description.journalClass | 1 | - |
dc.description.isOpenAccess | N | - |
dc.description.journalRegisteredClass | scopus | - |
dc.subject.keywordPlus | arginine | - |
dc.subject.keywordPlus | caspase 9 | - |
dc.subject.keywordPlus | cell penetrating peptide | - |
dc.subject.keywordPlus | cysteine | - |
dc.subject.keywordPlus | genomic DNA | - |
dc.subject.keywordPlus | glycine | - |
dc.subject.keywordPlus | guide RNA | - |
dc.subject.keywordPlus | leucine | - |
dc.subject.keywordPlus | polyhistidine tag | - |
dc.subject.keywordPlus | recombinant protein | - |
dc.subject.keywordPlus | virus vector | - |
dc.subject.keywordPlus | bacterial protein | - |
dc.subject.keywordPlus | Cas9 endonuclease Streptococcus pyogenes | - |
dc.subject.keywordPlus | cell penetrating peptide | - |
dc.subject.keywordPlus | endonuclease | - |
dc.subject.keywordPlus | guide RNA | - |
dc.subject.keywordPlus | amino terminal sequence | - |
dc.subject.keywordPlus | cell membrane | - |
dc.subject.keywordPlus | cell mutant | - |
dc.subject.keywordPlus | cytotoxicity | - |
dc.subject.keywordPlus | DNA sequence | - |
dc.subject.keywordPlus | expression vector | - |
dc.subject.keywordPlus | gene amplification | - |
dc.subject.keywordPlus | gene disruption | - |
dc.subject.keywordPlus | gene mutation | - |
dc.subject.keywordPlus | human | - |
dc.subject.keywordPlus | human cell | - |
dc.subject.keywordPlus | immune response | - |
dc.subject.keywordPlus | molecular cloning | - |
dc.subject.keywordPlus | nuclear localization signal | - |
dc.subject.keywordPlus | plasmid | - |
dc.subject.keywordPlus | polyacrylamide gel electrophoresis | - |
dc.subject.keywordPlus | promoter region | - |
dc.subject.keywordPlus | protein expression | - |
dc.subject.keywordPlus | RNA editing | - |
dc.subject.keywordPlus | RNA synthesis | - |
dc.subject.keywordPlus | bacterium transformation | - |
dc.subject.keywordPlus | chemistry | - |
dc.subject.keywordPlus | CRISPR Cas system | - |
dc.subject.keywordPlus | Escherichia coli | - |
dc.subject.keywordPlus | gene editing | - |
dc.subject.keywordPlus | genetics | - |
dc.subject.keywordPlus | HEK293 cell line | - |
dc.subject.keywordPlus | metabolism | - |
dc.subject.keywordPlus | procedures | - |
dc.subject.keywordPlus | Bacterial Proteins | - |
dc.subject.keywordPlus | Cell-Penetrating Peptides | - |
dc.subject.keywordPlus | Cloning, Molecular | - |
dc.subject.keywordPlus | CRISPR-Cas Systems | - |
dc.subject.keywordPlus | Endonucleases | - |
dc.subject.keywordPlus | Escherichia coli | - |
dc.subject.keywordPlus | Gene Editing | - |
dc.subject.keywordPlus | HEK293 Cells | - |
dc.subject.keywordPlus | Humans | - |
dc.subject.keywordPlus | Plasmids | - |
dc.subject.keywordPlus | RNA, Guide | - |
dc.subject.keywordPlus | Transformation, Bacterial | - |
dc.subject.keywordAuthor | Cas9 conjugation | - |
dc.subject.keywordAuthor | Cas9 protein purification | - |
dc.subject.keywordAuthor | Dialysis | - |
dc.subject.keywordAuthor | In vitro sgRNA synthesis | - |
dc.subject.keywordAuthor | Protein delivery | - |
dc.subject.keywordAuthor | T7E1 assay | - |
dc.identifier.url | https://link.springer.com/protocol/10.1007%2F978-1-4939-6518-2_7 | - |
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