0 439

Cited 0 times in

Cell-Penetrating Peptide-Mediated Delivery of Cas9 Protein and Guide RNA for Genome Editing

DC Field Value Language
dc.contributor.author김형범-
dc.contributor.author호사발레랑가파바라티-
dc.date.accessioned2017-11-02T08:10:35Z-
dc.date.available2017-11-02T08:10:35Z-
dc.date.issued2017-
dc.identifier.issn1064-3745-
dc.identifier.urihttps://ir.ymlib.yonsei.ac.kr/handle/22282913/154165-
dc.description.abstractThe 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. Cell-penetrating 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.-
dc.description.statementOfResponsibilityrestriction-
dc.languageEnglish-
dc.publisherHumana Press-
dc.relation.isPartOfMethods in Molecular Biology (Clifton, N.J.)-
dc.rightsCC BY-NC-ND 2.0 KR-
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/2.0/kr/-
dc.subject.MESHBacterial Proteins/genetics-
dc.subject.MESHBacterial Proteins/metabolism*-
dc.subject.MESHCRISPR-Cas Systems*-
dc.subject.MESHCell-Penetrating Peptides/chemistry-
dc.subject.MESHCell-Penetrating Peptides/metabolism*-
dc.subject.MESHCloning, Molecular-
dc.subject.MESHEndonucleases/genetics-
dc.subject.MESHEndonucleases/metabolism*-
dc.subject.MESHEscherichia coli-
dc.subject.MESHGene Editing/methods*-
dc.subject.MESHHEK293 Cells-
dc.subject.MESHHumans-
dc.subject.MESHPlasmids-
dc.subject.MESHRNA, Guide/chemistry-
dc.subject.MESHRNA, Guide/genetics-
dc.subject.MESHRNA, Guide/metabolism*-
dc.subject.MESHTransformation, Bacterial-
dc.titleCell-Penetrating Peptide-Mediated Delivery of Cas9 Protein and Guide RNA for Genome Editing-
dc.typeArticle-
dc.publisher.locationUnited States-
dc.contributor.collegeCollege of Medicine-
dc.contributor.departmentDept. of Pharmacology-
dc.contributor.googleauthorBharathi Suresh-
dc.contributor.googleauthorSuresh Ramakrishna-
dc.contributor.googleauthorHyongbum Kim-
dc.identifier.doi10.1007/978-1-4939-6518-2_7-
dc.contributor.localIdA04705-
dc.contributor.localIdA01148-
dc.relation.journalcodeJ02226-
dc.identifier.pmid27832534-
dc.identifier.urlhttps://link.springer.com/protocol/10.1007%2F978-1-4939-6518-2_7-
dc.subject.keywordCas9 conjugation-
dc.subject.keywordCas9 protein purification-
dc.subject.keywordDialysis-
dc.subject.keywordIn vitro sgRNA synthesis-
dc.subject.keywordProtein delivery-
dc.subject.keywordT7E1 assay-
dc.contributor.alternativeNameKim, Hyongbum-
dc.contributor.alternativeNameRamakrishna, S-
dc.contributor.affiliatedAuthorRamakrishna, S-
dc.contributor.affiliatedAuthorKim, Hyongbum-
dc.citation.titleMethods in Molecular Biology (Clifton, N.J.)-
dc.citation.volume1507-
dc.citation.startPage81-
dc.citation.endPage94-
dc.identifier.bibliographicCitationMethods in Molecular Biology (Clifton, N.J.), Vol.1507 : 81-94, 2017-
dc.date.modified2017-11-01-
dc.identifier.rimsid42127-
dc.type.rimsART-
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실) > 1. Journal Papers

qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.