Highly efficient Cas9-mediated transcriptional programming

Alejandro Chavez; Jonathan Scheiman; Suhani Vora; Benjamin W. Pruitt; Marcelle Tuttle; Eswar P R Iyer; Shuailiang Lin; Samira Kiani; Christopher D. Guzman; Daniel J. Wiegand; Dmitry Ter-Ovanesyan; Jonathan L. Braff; Noah Davidsohn; Benjamin E. Housden; Norbert Perrimon; Ron Weiss; John Aach; James J. Collins; George M. Church

doi:10.1038/nmeth.3312

Highly efficient Cas9-mediated transcriptional programming

Alejandro Chavez, Jonathan Scheiman, Suhani Vora, Benjamin W. Pruitt, Marcelle Tuttle, Eswar P R Iyer, Shuailiang Lin, Samira Kiani, Christopher D. Guzman, Daniel J. Wiegand, Dmitry Ter-Ovanesyan, Jonathan L. Braff, Noah Davidsohn, Benjamin E. Housden, Norbert Perrimon, Ron Weiss, John Aach, James J. Collins, George M. Church

Research output: Contribution to journal › Article › peer-review

1059 Scopus citations

Abstract

The RNA-guided nuclease Cas9 can be reengineered as a programmable transcription factor. However, modest levels of gene activation have limited potential applications. We describe an improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. We demonstrate its utility in activating endogenous coding and noncoding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human induced pluripotent stem cells (iPSCs).

Original language	English (US)
Pages (from-to)	326-328
Number of pages	3
Journal	Nature Methods
Volume	12
Issue number	4
DOIs	https://doi.org/10.1038/nmeth.3312
State	Published - Mar 31 2015
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

Access to Document

10.1038/nmeth.3312

Cite this

Chavez, A., Scheiman, J., Vora, S., Pruitt, B. W., Tuttle, M., P R Iyer, E., Lin, S., Kiani, S., Guzman, C. D., Wiegand, D. J., Ter-Ovanesyan, D., Braff, J. L., Davidsohn, N., Housden, B. E., Perrimon, N., Weiss, R., Aach, J., Collins, J. J., & Church, G. M. (2015). Highly efficient Cas9-mediated transcriptional programming. Nature Methods, 12(4), 326-328. https://doi.org/10.1038/nmeth.3312

Chavez, A, Scheiman, J, Vora, S, Pruitt, BW, Tuttle, M, P R Iyer, E, Lin, S, Kiani, S, Guzman, CD, Wiegand, DJ, Ter-Ovanesyan, D, Braff, JL, Davidsohn, N, Housden, BE, Perrimon, N, Weiss, R, Aach, J, Collins, JJ & Church, GM 2015, 'Highly efficient Cas9-mediated transcriptional programming', Nature Methods, vol. 12, no. 4, pp. 326-328. https://doi.org/10.1038/nmeth.3312

@article{bdc8c83ea2534bc99ca48427bd9b741e,

title = "Highly efficient Cas9-mediated transcriptional programming",

abstract = "The RNA-guided nuclease Cas9 can be reengineered as a programmable transcription factor. However, modest levels of gene activation have limited potential applications. We describe an improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. We demonstrate its utility in activating endogenous coding and noncoding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human induced pluripotent stem cells (iPSCs).",

author = "Alejandro Chavez and Jonathan Scheiman and Suhani Vora and Pruitt, {Benjamin W.} and Marcelle Tuttle and {P R Iyer}, Eswar and Shuailiang Lin and Samira Kiani and Guzman, {Christopher D.} and Wiegand, {Daniel J.} and Dmitry Ter-Ovanesyan and Braff, {Jonathan L.} and Noah Davidsohn and Housden, {Benjamin E.} and Norbert Perrimon and Ron Weiss and John Aach and Collins, {James J.} and Church, {George M.}",

note = "Funding Information: We thank K. Esvelt, P. Mali and the rest of the members of the Church and Collins labs for helpful discussions. We thank T. Ferrante, S. Byrne and M. Farrell for technical assistance, A. Keung (Massachusetts Institute of Technology) for providing zinc-finger reporter constructs, and J. Schulak for graphic design. This work was supported by US National Institutes of Health National Human Genome Research Institute grant P50 HG005550, US Department of Energy grant DE-FG02-02ER63445 and the Wyss Institute for Biologically Inspired Engineering. A.C. acknowledges funding by the National Cancer Institute grant 5T32CA009216-34. S.V. acknowledges funding by the National Science Foundation Graduate Research Fellowship Program, the Department of Biological Engineering at the Massachusetts Institute of Technology and the Department of Genetics at Harvard Medical School.",

year = "2015",

month = mar,

day = "31",

doi = "10.1038/nmeth.3312",

language = "English (US)",

volume = "12",

pages = "326--328",

journal = "Nature Methods",

issn = "1548-7091",

publisher = "Nature Publishing Group",

number = "4",

}

TY - JOUR

T1 - Highly efficient Cas9-mediated transcriptional programming

AU - Chavez, Alejandro

AU - Scheiman, Jonathan

AU - Vora, Suhani

AU - Pruitt, Benjamin W.

AU - Tuttle, Marcelle

AU - P R Iyer, Eswar

AU - Lin, Shuailiang

AU - Kiani, Samira

AU - Guzman, Christopher D.

AU - Wiegand, Daniel J.

AU - Ter-Ovanesyan, Dmitry

AU - Braff, Jonathan L.

AU - Davidsohn, Noah

AU - Housden, Benjamin E.

AU - Perrimon, Norbert

AU - Weiss, Ron

AU - Aach, John

AU - Collins, James J.

AU - Church, George M.

N1 - Funding Information: We thank K. Esvelt, P. Mali and the rest of the members of the Church and Collins labs for helpful discussions. We thank T. Ferrante, S. Byrne and M. Farrell for technical assistance, A. Keung (Massachusetts Institute of Technology) for providing zinc-finger reporter constructs, and J. Schulak for graphic design. This work was supported by US National Institutes of Health National Human Genome Research Institute grant P50 HG005550, US Department of Energy grant DE-FG02-02ER63445 and the Wyss Institute for Biologically Inspired Engineering. A.C. acknowledges funding by the National Cancer Institute grant 5T32CA009216-34. S.V. acknowledges funding by the National Science Foundation Graduate Research Fellowship Program, the Department of Biological Engineering at the Massachusetts Institute of Technology and the Department of Genetics at Harvard Medical School.

PY - 2015/3/31

Y1 - 2015/3/31

N2 - The RNA-guided nuclease Cas9 can be reengineered as a programmable transcription factor. However, modest levels of gene activation have limited potential applications. We describe an improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. We demonstrate its utility in activating endogenous coding and noncoding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human induced pluripotent stem cells (iPSCs).

AB - The RNA-guided nuclease Cas9 can be reengineered as a programmable transcription factor. However, modest levels of gene activation have limited potential applications. We describe an improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. We demonstrate its utility in activating endogenous coding and noncoding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human induced pluripotent stem cells (iPSCs).

UR - http://www.scopus.com/inward/record.url?scp=84926521955&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84926521955&partnerID=8YFLogxK

U2 - 10.1038/nmeth.3312

DO - 10.1038/nmeth.3312

M3 - Article

C2 - 25730490

AN - SCOPUS:84926521955

SN - 1548-7091

VL - 12

SP - 326

EP - 328

JO - Nature Methods

JF - Nature Methods

IS - 4

ER -

Highly efficient Cas9-mediated transcriptional programming

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this