Wang et al., "TALE nickase
mediates high efficient targeted transgene integration at the human multicopy ribosomal DNA locus," Biochemical and Biophysical Research Communications, vol.
Kim et al., "Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases
," Genome Research, vol.
Singhal et al., "Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase
applications," Genome Biology, vol.
 Shen, B., et al., Efficient genome modification by CRISPR-Cas9 nickase
with minimal off-target effects.
Plasmid Construction and Gene Targeting Mediated by CRISPR/Cas9 Nickase
. To create the donor vector, cassettes consisting of marker genes NeoR, EGFP, and HSVtk and inverted loxP sequences were constructed into the EcoRI-BamHI site of pBluescript SK.
The CRISPR/ Cas system can be used as nuclease for in planta gene targeting and as paired nickases
for directed mutagenesis in Arabidopsis resulting in heritable progeny.
The Cas9D10A, mutant CRISPR Nickase
, has been produced to selectively make a single-strand DNA cut at the target sequence.
, which is a mutated Cas9 (Cas9-D10A) enzyme, was newly developed for precise genomic modification .
variants (Cas9-D10A or Cas9-H840A) containing a single inactive nuclease domain cleave only one DNA strand to create a single-strand break at the target sites.
The molecular biology grade ethanol has no detectable Dnase, Rnase or Nickase