title: RNA-guided gene activation by CRISPR-Cas9-based transcription factors.
abstract: Technologies for engineering synthetic transcription factors have enabled many advances in medical and scientific research. In contrast to existing methods based on engineering of DNA-binding proteins, we created a Cas9-based transactivator that is targeted to DNA sequences by guide RNA molecules. Coexpression of this transactivator and combinations of guide RNAs in human cells induced specific expression of endogenous target genes, demonstrating a simple and versatile approach for RNA-guided gene activation.
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA.
authors.forename: Pablo
authors.initials: P
authors.lastname: Perez-Pinera
authors.forename: D Dewran
authors.initials: DD
authors.lastname: Kocak
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Vockley
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Adler
authors.forename: Ami M
authors.initials: AM
authors.lastname: Kabadi
authors.forename: Lauren R
authors.initials: LR
authors.lastname: Polstein
authors.forename: Pratiksha I
authors.initials: PI
authors.lastname: Thakore
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Glass
authors.forename: David G
authors.initials: DG
authors.lastname: Ousterout
authors.forename: Kam W
authors.initials: KW
authors.lastname: Leong
authors.forename: Farshid
authors.identifier: 0000000173800330
authors.initials: F
authors.lastname: Guilak
authors.forename: Gregory E
authors.initials: GE
authors.lastname: Crawford
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Reddy
authors.forename: Charles A
authors.initials: CA
authors.lastname: Gersbach
mesh_terms: D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D017393:RNA Editing; D012260:Ribonucleases / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*; D015533:Transcriptional Activation; D058727:RNA, Small Untranslated
chemical_list: C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D014157:Transcription Factors; D012260:Ribonucleases; D058727:RNA, Small Untranslated
references.citation: Mol Ther Nucleic Acids. 2012 Jul 10;1:e32
references.pmid: 23344148
references.citation: Nat Med. 2002 Dec;8(12):1427-32
references.pmid: 12415262
references.citation: Nat Methods. 2013 Mar;10(3):243-5
references.pmid: 23396285
references.citation: Nat Methods. 2013 Mar;10(3):239-42
references.pmid: 23377379
references.citation: Nucleic Acids Res. 2012 Nov;40(21):10596-613
references.pmid: 23002135
references.citation: Mol Cell. 2011 Sep 16;43(6):915-26
references.pmid: 21925380
references.citation: Oncogene. 2007 Apr 26;26(19):2791-8
references.pmid: 17057734
references.citation: Nat Biotechnol. 2011 Feb;29(2):149-53
references.pmid: 21248753
references.citation: Stem Cells. 2003;21(6):632-7
references.pmid: 14595122
references.citation: Genome Biol. 2010;11(10):R106
references.pmid: 20979621
references.citation: Nucleic Acids Res. 2012 Aug;40(15):7584-95
references.pmid: 22581776
references.citation: Science. 2013 Feb 15;339(6121):819-23
references.pmid: 23287718
references.citation: Genome Biol. 2009;10(3):R25
references.pmid: 19261174
references.citation: Nat Biotechnol. 2003 Mar;21(3):269-74
references.pmid: 12592412
references.citation: Nucleic Acids Res. 2012 Jul;40(12):5368-77
references.pmid: 22387464
references.citation: Nat Biotechnol. 2011 Feb;29(2):143-8
references.pmid: 21179091
references.citation: Nat Biotechnol. 2002 Feb;20(2):135-41
references.pmid: 11821858
references.citation: Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1495-500
references.pmid: 10660690
references.citation: Nat Biotechnol. 2003 Oct;21(10):1208-14
references.pmid: 12960965
references.citation: J Biol Chem. 2005 Feb 4;280(5):3707-14
references.pmid: 15537646
references.citation: Nat Biotechnol. 2013 Mar;31(3):230-2
references.pmid: 23360966
references.citation: Sci Rep. 2012;2:897
references.pmid: 23193439
references.citation: Cell. 2013 Feb 28;152(5):1173-83
references.pmid: 23452860
references.citation: Nucleic Acids Res. 2012 Jun;40(11):5180-7
references.pmid: 22323524
references.citation: Science. 2012 Aug 17;337(6096):816-21
references.pmid: 22745249
references.citation: Science. 2013 Feb 15;339(6121):823-6
references.pmid: 23287722
references.citation: Genome Res. 2012 Jan;22(1):134-41
references.pmid: 22128135
references.citation: Nature. 2010 Feb 25;463(7284):1035-41
references.pmid: 20107439
references.citation: Elife. 2013 Jan 29;2:e00471
references.pmid: 23386978
references.citation: Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14628-33
references.pmid: 9843940