CRISPR/Cas9 - possibilities in the future
CRISPR has been a popular topic of late and has invoked interest in countless people in recent times.
Clustered regularly interspaced short palindromic repeats (CRISPR, pronounced crisper) are segments of prokaryotic DNA containing short repetitions of base sequences. Each repetition is followed by short segments of "spacer DNA" from previous exposures to a bacteriophage virus or plasmid.
Cas9 (CRISPR associated protein 9) is an RNA-guided DNA endonuclease enzyme associated with the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) adaptive immunity system in Streptococcus pyogenes, among other bacteria. S. pyogenes utilizes Cas9 to memorize and later interrogate and cleave foreign DNA, such as invading bacteriophage DNA or plasmid DNA.
For the first time in 2012, Virginijus Šikšnys together with G. Gašiūnas, R. Barrangou, and P. Horvath, purified Cas9 in complex with crRNA from the E. coli strain engineered to carry the S. thermophilus CRISPR locus and undertook a series of biochemical experiments to mechanistically characterize Cas9’s mode of action.
Following its initial demonstration in 2012, the CRISPR/Cas9 system has been widely adopted. This has already been successfully used to target important genes in many cell lines and organisms, including human, bacteria, zebrafish, C. elegans, plants, Xenopus tropicalis, yeast, Drosophila, monkeys, rabbits, pigs, rats and mice.
Several groups have now taken advantage of this method to introduce single point mutations (deletions or insertions) in a particular target gene, via a single gRNA.
Using a pair of gRNA-directed Cas9 nucleases instead, it is also possible to induce large deletions or genomic rearrangements, such as inversions or translocations. A recent exciting development is the use of the dCas9 version of the CRISPR/Cas9 system to target protein domains for transcriptional regulation,epigenetic modification, and microscopic visualization of specific genome loci.
For the first time, scientists have edited DNA in healthy and viable human embryos using genetic tool CRIPR/Cas9. The researchers, led by developmental biologist Fredrik Lanner from the Karolinska Institutet in Sweden, hope the research will lead to new ways to treat infertility and prevent miscarriage.
The rapid progress in developing Cas9 into a set of tools for cell and molecular biology research has been remarkable, likely due to the simplicity, high efficiency and versatility of the system. Of the designer nuclease systems currently available for precision genome engineering, the CRISPR/Cas system is by far the most user friendly. It is now also clear that Cas9’s potential reaches beyond DNA cleavage, and its usefulness for genome locus-specific recruitment of proteins will likely only be limited by our imagination.
Check out future posts for updates on CRISPR!
Clustered regularly interspaced short palindromic repeats (CRISPR, pronounced crisper) are segments of prokaryotic DNA containing short repetitions of base sequences. Each repetition is followed by short segments of "spacer DNA" from previous exposures to a bacteriophage virus or plasmid.
Cas9 (CRISPR associated protein 9) is an RNA-guided DNA endonuclease enzyme associated with the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) adaptive immunity system in Streptococcus pyogenes, among other bacteria. S. pyogenes utilizes Cas9 to memorize and later interrogate and cleave foreign DNA, such as invading bacteriophage DNA or plasmid DNA.
For the first time in 2012, Virginijus Šikšnys together with G. Gašiūnas, R. Barrangou, and P. Horvath, purified Cas9 in complex with crRNA from the E. coli strain engineered to carry the S. thermophilus CRISPR locus and undertook a series of biochemical experiments to mechanistically characterize Cas9’s mode of action.
Following its initial demonstration in 2012, the CRISPR/Cas9 system has been widely adopted. This has already been successfully used to target important genes in many cell lines and organisms, including human, bacteria, zebrafish, C. elegans, plants, Xenopus tropicalis, yeast, Drosophila, monkeys, rabbits, pigs, rats and mice.
Several groups have now taken advantage of this method to introduce single point mutations (deletions or insertions) in a particular target gene, via a single gRNA.
Using a pair of gRNA-directed Cas9 nucleases instead, it is also possible to induce large deletions or genomic rearrangements, such as inversions or translocations. A recent exciting development is the use of the dCas9 version of the CRISPR/Cas9 system to target protein domains for transcriptional regulation,epigenetic modification, and microscopic visualization of specific genome loci.
For the first time, scientists have edited DNA in healthy and viable human embryos using genetic tool CRIPR/Cas9. The researchers, led by developmental biologist Fredrik Lanner from the Karolinska Institutet in Sweden, hope the research will lead to new ways to treat infertility and prevent miscarriage.
The rapid progress in developing Cas9 into a set of tools for cell and molecular biology research has been remarkable, likely due to the simplicity, high efficiency and versatility of the system. Of the designer nuclease systems currently available for precision genome engineering, the CRISPR/Cas system is by far the most user friendly. It is now also clear that Cas9’s potential reaches beyond DNA cleavage, and its usefulness for genome locus-specific recruitment of proteins will likely only be limited by our imagination.
Check out future posts for updates on CRISPR!
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