Re: [DIYbio] Editing the genome with high precision

[Nathan McCorkle]

Cool they're using CRISPRs! Looks pretty good, their website has a
reagent section which mentions addgene and plasmids. I found another
paper that was published the same day referencing it...

Multiplex Genome Engineering Using CRISPR/Cas Systems
http://www.sciencemag.org/content/early/2013/01/03/science.1231143
http://zlab.mit.edu/reprints/Cong_Science_2013.pdf
"
Functional elucidation of causal genetic variants and elements
requires precise genome editing technologies. The type II prokaryotic
CRISPR (clustered regularly interspaced short palindromic repeats)
adaptive immune system has been shown to facilitate RNA-guided
site-specific DNA cleavage. We engineered two different type II CRISPR
systems and demonstrate that Cas9 nucleases can be directed by short
RNAs to induce precise cleavage at endogenous genomic loci in human
and mouse cells. Cas9 can also be converted into a nicking enzyme to
facilitate homology-directed repair with minimal mutagenic activity.
Finally, multiple guide sequences can be encoded into a single CRISPR
array to enable simultaneous editing of several sites within the
mammalian genome, demonstrating easy programmability and wide
applicability of the CRISPR technology.
"

Reagents section
http://www.genome-engineering.org/crispr/?page_id=23

RNA-guided human genome engineering via Cas9
http://arep.med.harvard.edu/pdf/Mali_Sci_12.pdf
"
Bacteria and archaea have evolved adaptive immune defenses termed clustered
regularly interspaced short palindromic repeats
(CRISPR)/CRISPR-associated (Cas)
systems that use short RNA to direct degradation of foreign nucleic
acids. Here, we
engineer the type II bacterial CRISPR system to function with custom guide RNA
(gRNA) in human cells. For the endogenous AAVS1 locus, we obtained targeting
rates of 10 to 25% in 293T cells, 13 to 8% in K562 cells, and 2 to 4%
in induced
pluripotent stem cells. We show this process relies on CRISPR components, is
sequence-specific, and upon simultaneous introduction of multiple gRNAs, can
effect multiplex editing of target loci. We also compute a genome-wide
resource of
~190k unique gRNAs targeting ~40.5% of human exons. Our results establish an
RNA-guided editing tool for facile, robust, and multiplexable human genome
engineering.
"

On Sat, Jan 5, 2013 at 7:55 PM, Cory Geesaman <co...@geesaman.com> wrote:
> http://web.mit.edu/newsoffice/2013/editing-the-genome-with-high-precision-0103.html
>
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[John Griessen]

On 01/05/2013 09:55 PM, Cory Geesaman wrote:
> http://web.mit.edu/newsoffice/2013/editing-the-genome-with-high-precision-0103.html
>

"The research team has deposited the necessary genetic components with a nonprofit called Addgene, making the components widely
available to other researchers who want to use the system. The researchers have also created a website with tips and tools for
using this new technique." --> http://www.genome-engineering.org/


"Please check back soon, the plasmids are being processed by Addgene. We will post links as soon as Addgene finishes processing
our depositions. "

Seems like a breakthrough that is being purposely left open, unlocked, or am I fooling?

[Cathal Garvey]

If so, I dearly love these guys/girls already.

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