Research and publish the best content.
Get Started for FREE
Sign up with Facebook Sign up with X
I don't have a Facebook or a X account
Already have an account: Login
Genetic Engineering Publications - GEG Tech top picks
36.6K views |
+0 today
 Your new post is loading...
Your new post is loading...
Scoop.it!
BigField GEG Tech's insight:
In this report, CRISPR/Cas9 system was used to make large DNA fragment deletions and insertions, including entire Dip2a gene deletion, about 65kb in size, and β-galactosidase (lacZ) reporter gene insertion of larger than 5kb in mouse. About 11.8% (11/93) are positive for 65kb deletion from transfected and diluted ES clones. High targeting efficiencies in ES cells were also achieved with G418 selection, 46.2% (12/26) and 73.1% (19/26) for left and right arms respectively. Targeted large fragment deletion efficiency is about 21.4% of live pups or 6.0% of injected embryos. Targeted insertion of lacZ reporter with NEO cassette showed 27.1% (13/48) of targeting rate by ES cell transfection and 11.1% (2/18) by direct zygote injection. 
|
Scoop.it!
Profiling of engineering hotspots identifies an allosteric CRISPR-Cas9 switch - Nature Biotechnology
A small-molecule-inducible Cas9 variant with very low background activity is identified by screening for sites that can tolerate domain insertions.
BigField GEG Tech's insight:
Here the scientists experimentally profile Cas9 using randomized insertional mutagenesis and delineate hotspots in the structure capable of tolerating insertions of a PDZ domain without disruption of the enzyme's binding and cleavage functions. They find that domain insertion profiling facilitates the rapid generation of new Cas9 functionalities and provides useful data for future engineering of Cas9.
|
A team led by researchers from Broad Institute of MIT and Harvard, and the McGovern Institute for Brain Research at MIT, has characterized and engineered a new gene-editing system that can precisely and efficiently insert large DNA sequences into a genome.
Science Publication:
RNA-guided DNA insertion with CRISPR-associated transposases