Novel approach to gene regulation can activate multiple genes simultaneously

Daigram of enzyme used in CRISPR-on

Whitehead Institute researchers created the enzyme used in CRISPR-on by fusing the Cas9 protein to a VP160 domain containing 10 tandem copies of VP16 motifs.  The VP160 domain acts as a transcriptional activation domain.

Image: Courtesy of Cell Research

August 27, 2013

Tags: Jaenisch LabStem Cells + Therapeutic CloningGenetics + Genomics

CAMBRIDGE, Mass.  – By creating a powerful new gene regulation system called CRISPR-on, Whitehead Institute researchers now have the ability to increase the expression of multiple genes simultaneously and precisely manipulate each gene’s expression level. The system is effective in both mouse and human cells as well as in mouse embryos.

“CRISPR-on is a tool that will be very useful for studying many biological processes, particularly for studying gene functions and gene networks,” says Whitehead Founding Member Rudolf Jaenisch. “In contrast to RNA interference, which is commonly used to inactivate gene activity, the CRISPR-on system allows activation of cellular genes. The technology substantially expands our ability to change gene expression in cultured cells and animals..”

The system, called CRISPR-on, is a modified version of CRISPR/Cas (for “clustered regularly interspaced short palindromic repeat/CRISPR associated”), which taps into a bacterial defense system against viral intruders. CRISPR/Cas relies on an enzyme, Cas9, which cuts DNA at locations specified by single guide RNAs (sgRNAs). For CRISPR-on, the Whitehead team modified the Cas9 enzyme by eliminating its ability to cleave DNA and adding a transcription activation domain. The resulting enzyme can increase gene expression without permanently changing the DNA.

The new system is described this week in the journal Cell Research.

CRISPR-on’s ability to activate only the desired genes at varying levels could be used to help scientists improve our understanding of transcription network underlying a variety of diseases and potentially find new ways to treat them.

“Many diseases, especially complex diseases, involve multiple genes, and this system could be used therapeutically to target and activate multiple genes together and rescue these disease phenotypes,” says Albert Cheng, a graduate student in the Jaenisch lab and co-author of the Cell Research paper. “Or we could use it to study the gene networks in diseases and get a better understanding of how those diseases work.”

So far, the researchers have used CRISPR-on to activate up to three native genes concurrently in human cells.

“I think we need to do more work to see if there are any limitations to the number of genes CRISPR-on can activate at a time,” says Haoyi Wang, a co-author and postdoctoral researcher in the Jaenisch lab. “We’d like to see if we can get data on activating 10 or more genes, to see if there is an upper limit to what this system can do.”

 

This work is supported by the Croucher Foundation and National Institutes of Health (NIH) grants HD 045022 and R37CA084198.

 

Written by Nicole Giese Rura

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Rudolf Jaenisch's primary affiliation is with Whitehead Institute for Biomedical Research, where his laboratory is located and all his research is conducted. He is also a professor of biology at Massachusetts Institute of Technology.

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Full Citation:

“Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system”

Cell Research, online August 27, 2013.

Albert W. Cheng (1,2,5), Haoyi Wang (1,5), Hui Yang (1), Linyu Shi (1), Yarden Katz (1,3), Sudharshan Rangarajan (1), Thorold W. Theunissen (1), Chikdu S. Shivalila (1,4), Daniel B. Dadon (1,4), Rudolf Jaenisch (1,4).

1. Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA
2. Computational and Systems Biology Program, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA
3. Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA
4. Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA
5. These author contributed equally

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