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Two alternative approaches gave more promising results: the use of Cas9 nickase (Cas9n) or fCas9.
An update on the tools for creating transgenic animal models of human diseases--focus on atherosclerosis
Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects.
Deactivated CRISPR Associated Protein 9 for Minor-Allele Enrichment in Cell-Free DNA
The TadA A106V/D108N mutant was fused to the N-terminus of the Cas9 (D10A mutant with nickase activity) [52].
Protein Engineering Strategies to Expand CRISPR-Cas9 Applications
deactivated the catalytic domain of the nuclease creating a nickase [75].
Genome Editing Redefines Precision Medicine in the Cardiovascular Field
Though, increasing specificity by titrating Cas9-sgRNA concentration also reduces on-target frequencies [43]; (b) wild-type cas9 is mutated to D10 mutant nickase and paired with two sgRNAs to induce nick in the opposite strands of two nearby target sites.
CRISPR/Cas9 System: A Bacterial Tailor for Genomic Engineering
This strategy known as nickase Cas9 can be achieved by inactivation of one of the two nuclease domains of the Cas9, resulting in the cleavage of only one DNA strand.
Guiding Lights in Genome Editing for Inherited Retinal Disorders: Implications for Gene and Cell Therapy
Zhang et al., "Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects," Nature Methods, vol.
Aberrant Expression of miR-362 Promotes Lung Cancer Metastasis through Downregulation of Sema3A
Single nucleotide mutation in two domains (RuvC, HNH) of Cas9 transforms it into Cas9 nickase [5, 9].
CRISPR CAS9 SYSTEM: A NOVEL GENOME EDITING TOOL
This cassette was integrated into the target site by homologous recombination using the clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins 9 (CRISPR/Cas9) nickase system [6-8].
Engineering of Systematic Elimination of a Targeted Chromosome in Human Cells
Cas9 specificity was increase 100-1500 fold by the use of a pair of Cas9 nickase variants, which were directed to target on opposite strands up to 100 bp apart.
RNA-guided genome editing tool CRISPR-Cas9: its applications and achievements in model and crop plants
The company offers not just plasmids for CRISPR gene editing, but also custom-guide RNA design and validation (to provide customers with confidence that they are working with a suitably active guide); custom donor design and manufacturing; QuickStart cell lines that constitutively produce Cas9 nickase; and more than 550 X-MAN cell lines ready for further modification.
Kits make genome editing accessible to everyone
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