What is CRISPR-Cas9?
Working with DNA is a common base for almost all life science branches. The manipulation of genes is often the method of choice to get insight into cellular processes. Thus DNA-editing proteins which are able to bind and open DNA strands site-specifically are of great importance. There are several to choose from, and all have their advantages and disadvantages. The Cas9 endonuclease of the bacterial derived CRISPR-Cas9 immune system is one of them. In contrast to its alternatives – meganucleases, ZFNs, and TALENs – the Cas9 DNA-binding determinant is an RNA molecule, which is easy to design. Thus Cas9 is very flexible concerning target sequence choice. Therefore it is frequently used to mutate DNA, and to produce knock-out and knock-in mutants.
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Role of circadian gene Clock during differentiation of mouse pluripotent stem cells
Biological rhythms controlled by the circadian clock are absent in embryonic stem cells (ESCs). However, they start to develop during the differentiation of pluripotent ESCs to downstream cells. Conversely, biological rhythms in adult somatic cells disappear when they are reprogrammed into induced pluripotent stem cells (iPSCs). These studies indicated that the development of biological rhythms in ESCs might be closely associated with the maintenance and differentiation of ESCs.Read article -
Video: Genome Engineering with CRISPR-Cas9
Jennifer Doudna tells the story of how studying the way bacteria fight viral infection turned into a genomic engineering technology that has transformed molecular biology research. In 2013, Doudna and her colleagues developed the CRISPR-Cas9 gene expression system that, when introduced into animal cells, makes site-specific changes to intact genomes. CRISPR-Cas9 is more precise, more efficient, and less expensive than other genome editing tools and, as a result, has facilitated a wide range of studies that were previously unachievable.Read article -
Video: Genome Editing with CRISPR-Cas9
Jennifer Doudna tells the story of how studying the way bacteria fight viral infection turned into a genomic engineering technology that has transformed molecular biology research. In 2013, Doudna and her colleagues developed the CRISPR-Cas9 gene expression system that, when introduced into animal cells, makes site-specific changes to intact genomes. CRISPR-Cas9 is more precise, more efficient, and less expensive than other genome editing tools and, as a result, has facilitated a wide range of studies that were previously unachievable.Read article -
Five Big Mysteries about CRISPR’s Origins
Francisco Mojica was not the first to see CRISPR, but he was probably the first to be smitten by it. He remembers the day in 1992 when he got his first glimpse of the microbial immune system that would launch a biotechnology revolution. He was reviewing genome-sequence data from the salt-loving microbe Haloferax mediterranei and noticed 14 unusual DNA sequences, each 30 bases long.Read article -
Gene Editing with CRISPR/Cas9 - Breakthrough in Genome Engineering
The CRISPR/Cas9 system is one of several different bacterial systems for defense against viral attacks. It consists of two main components. One is a small piece of RNA which binds to the viral target sequence via Watson-Crick base pairing. It serves as a marker for the foreign nucleic acid. The second component is the Cas9 protein. It binds to the marked sequence and cuts it due to its nuclease activity. Because the base pairing RNA can be synthesized easily and then used to determine a target region, researchers have utilized this system in the laboratory for genome editing.Read article -
Video Talk: CRISPR-Cas – From a Bacterial Adaptive Immune System to a Genome Engineering Tool
The CRISPR-Cas system was originally discovered as an adaptive immune system of bacteria and archaea to protect against viral attack. During this talk, given at Leica Microsystems in Wetzlar, Dr. Lennart Randau, MPI Marburg, presents this simple and fascinating system in detail. Furthermore, he introduces the adaption of the CRISPR-Cas system into a potent molecular biology tool, which is used heavily for genome editing. In addition to its influence on molecular biology, meanwhile the Cas9 nuclease also stimulates microscopy techniques e.g. to fluorescently label genomic loci in living cells.Read article -
CRISPR/Cas9-mediated Endogenous Protein Tagging for RESOLFT Super-Resolution Microscopy of Living Human Cells
Overexpression is a notorious concern in conventional and especially in super-resolution fluorescence light microscopy studies because it may cause numerous artifacts including ectopic sub-cellular localizations, erroneous formation of protein complexes, and others. Nonetheless, current live cell super-resolution microscopy studies generally rely on the overexpression of a host protein fused to a fluorescent protein.Read article