Highly efficient Cas9-mediated transcriptional programming.
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).
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, USA. [3] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Alejandro
A
Chavez
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Jonathan
J
Scheiman
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. [3] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Suhani
S
Vora
Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.
Benjamin W
BW
Pruitt
Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.
Marcelle
M
Tuttle
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Eswar
E
P R Iyer
1] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA. [3] School of Life Sciences, Tsinghua University, Beijing, China.
Shuailiang
S
Lin
1] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [2] Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Samira
S
Kiani
Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.
Christopher D
CD
Guzman
Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.
Daniel J
DJ
Wiegand
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Dmitry
D
Ter-Ovanesyan
Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.
Jonathan L
JL
Braff
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Noah
N
Davidsohn
1] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.
Benjamin E
BE
Housden
1] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.
Norbert
N
Perrimon
1] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [2] Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [3] Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Ron
R
Weiss
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
John
J
Aach
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [3] Institute for Medical Engineering &Science, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [4] Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
James J
JJ
Collins
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
George M
GM
Church
D002454:Cell Differentiation / Q000235:genetics; D004720:Endonucleases / Q000235:genetics; D005821:Genetic Techniques; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D009474:Neurons / Q000166:cytology; D013211:Staphylococcus aureus; D015533:Transcriptional Activation; D000094704:RNA, Guide, CRISPR-Cas Systems
D004720:Endonucleases; D000094704:RNA, Guide, CRISPR-Cas Systems