On March 7th, the website of the journal Nature published an article entitled article "CRISPR gene editing is just the beginning", pointing out that gene editing is only a starting point of CRISPR technology application. The real ability of this biological tool in these feature stories in Nature is to explore the operational mechanism of the genome. And it introduced technological achievements and applications published on Nature, Science, Cell and their sub-journals recently.
Epigenetic modification on DNA is not the only genome code to be cracked. Over 98% of the human genome does not encode proteins. But the researchers believe that a considerable part of such DNA doing important things, they are adopting CRISPR-Cas9 to clarify what it is.
Some DNA encoding such as microRNAs and long non-coding RNA molecule of a class - people think they have a function other than protein synthesis. Other sequences can be required to enlarge the gene expression "enhancers." Most common risks associated with the DNA sequence located in the genomic region containing the non-coding RNA and enhancers in. But before CRISPR-Cas9, researchers were difficult to figure out the function of these sequences. Boston Children's Hospital hematologist Daniel Bauer said, "We do not have a good way of these non-coding genomic annotation from the feature is now more sophisticated in our experiments."
University of Southern California molecular biologist Peggy Farnham and colleagues used CRISPR-Cas9 delete mutations in prostate cancer and colorectal cancer genome research found enhancer region. Sometimes the findings surprised her. In an experiment not released, her team deleted a past that is important enhancer, not its one million bases in a gene expression changes. "It happened when we usually classify a regulatory element influence the way you remove the element and does not match."
When possible use of CRISPR-Cas9 researchers to explore the large fragment of DNA regulation will be more surprises. From the Massachusetts Institute of Technology (MIT) genetic David Gifford and Brigham and Women's Hospital research team led by Richard Sherwood advantage of this technology in the base sequence of some 40,000 manufacturing mutation, then examined whether each change would near fluorescent protein coding genes affected. Thereby resulting in a DNA sequence to enhance gene expression profiles, including several based on characteristics such as gene regulation chromatin modification is not predicted sequence (CRISPR NEW, Looking for gene regulatory elements, High-throughput mapping of regulatory DNA.Nature Biotechnology 34, 167-174 (2016) doi: 10.1038 / nbt.3468).
In-depth study of this dark matter is faced with challenges even using CRISPR-Cas9. Cas9 wizard RNA cleavage enzyme will inform its site, but only if there is a specific and near the common DNA sequence cleavage site. Researchers who want to silence the gene is not difficult, because these important sequences are almost always exists somewhere within the gene. But for those who want to complete a short non-coding RNAs are very specific changes for the limited choice. Netherlands Cancer Institute researcher Reuven Agami, said, "We cannot get any sequence."
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