@article {44489, title = {G-quadruplex motif at the 3{\textquoteright} end of sgRNAs improves CRISPR-Cas9 based genome editing efficiency}, journal = {Chemical Communications}, volume = {54}, year = {2018}, month = {MAR}, pages = {2377-2380}, abstract = {Originating as a component of prokaryotic adaptive immunity, the type II CRISPR/Cas9 system has been repurposed for targeted genome editing in various organisms. Although Cas9 can bind and cleave DNA efficiently under in vitro conditions, its activity inside a cell can vary dramatically between targets owing to differences between genomic loci and availability of enough Cas9/sgRNA (single guide RNA) complex molecules for cleavage. Most methods to improve CRISPR/Cas9 activity have so far relied on Cas9 protein engineering or base modifications in the sgRNA sequence. Here we demonstrate that a structure based rational design of sgRNAs can enhance the efficiency of Cas9 cleavage in vivo. By appending a naturally forming RNA G-quadruplex motif to the 3' end of sgRNAs we can improve its stability and target cleavage efficiency in zebrafish embryos without inducing off-target activity, thereby underscoring its value in the design of better and optimized genome editing triggers.}, issn = {1359-7345}, doi = {10.1039/C7CC08893K}, author = {Nahar, Smita and Sehgal, Paras and Azhar, Mohammad and Rai, Manish and Singh, Amrita and Sivasubbu, Sridhar and Chakraborty, Debojyoti and Maiti, Souvik} }