<p>Target sites and alleles are shown for four genes for pools of embryos or single embryos (n = 12 per target with Cas9 mRNA, n = 12 per target with Cas9 protein). For each target site, the top panel is from a pool of embryos while the bottom panel represents alleles from single embryos. Each plot indicates the observed mutant alleles using arches that connect the bases surrounding the deletion. In the top panel, the y-axis and color of the arch indicate allele abundance; for ease of visualization, the bottom panel indicates allele abundance only with color. PAM is indicated on the DNA sequence in uppercase. g<i>ria3a</i> sgRNA induces only low indel frequencies when injected with Cas9 mRNA.</p
(A) Coding sequence of CASP9 targeted by sgRNA3 used in this work. The CASP9-sgRNA-specific region (...
<p>(A) Schematic showing the recombinant adenoviral genome-harbored <i>EGFP</i> (ADV-EGFP), with the...
The Streptococcus pyogenes Cas9 (SpCas9) nuclease can be efficiently targeted to genomic loci by mea...
<p>Data in panels A, C, E, G from microinjection of sgRNA chr17(+):34030557–34030576 and panels B, D...
<p><b>A.</b> Diagram of Cas9/sgRNA complex injection. <b>B.</b> Purified His-tagged Cas9 protein com...
Understanding the impact of guide RNA (gRNA) and genomic locus on CRISPR-Cas9 activity is crucial to...
CRISPR/Cas9 systems enable many molecular activities to be efficiently directed in vivo to user-spec...
<p>(A) Schematic illustration of the RGN system. Engineered sgRNA:Cas9 system is depicted here based...
a.Cas9_1: Mali et al., 2013 [3]. Cas9_2: Fauser et al., 2014 [13]. Cas9_3: Li et al., 2013 [25]. Cas...
Here, we present a generalized method of guide RNA “tuning” that enables Cas9 to discriminate betwee...
<p>(A) A schematic diagram showing primers for detecting mutations in Cas9 target site of <i>nsd2</i...
(A) Schematic depicting Cas9 variants and orientation of sgRNAs required to generate blunt, 5’, and ...
<p>a) An in vitro digestion assay shows that sgRNAs differ in their efficiencies. Below the gene nam...
<p>A) Percentage mutation rates (mutant embryos per total number) for maternal Cas9 supply (heterozy...
We report that engineered Cas9 variants with improved specificity-eCas9-1.1 and Cas9-HF1-are often p...
(A) Coding sequence of CASP9 targeted by sgRNA3 used in this work. The CASP9-sgRNA-specific region (...
<p>(A) Schematic showing the recombinant adenoviral genome-harbored <i>EGFP</i> (ADV-EGFP), with the...
The Streptococcus pyogenes Cas9 (SpCas9) nuclease can be efficiently targeted to genomic loci by mea...
<p>Data in panels A, C, E, G from microinjection of sgRNA chr17(+):34030557–34030576 and panels B, D...
<p><b>A.</b> Diagram of Cas9/sgRNA complex injection. <b>B.</b> Purified His-tagged Cas9 protein com...
Understanding the impact of guide RNA (gRNA) and genomic locus on CRISPR-Cas9 activity is crucial to...
CRISPR/Cas9 systems enable many molecular activities to be efficiently directed in vivo to user-spec...
<p>(A) Schematic illustration of the RGN system. Engineered sgRNA:Cas9 system is depicted here based...
a.Cas9_1: Mali et al., 2013 [3]. Cas9_2: Fauser et al., 2014 [13]. Cas9_3: Li et al., 2013 [25]. Cas...
Here, we present a generalized method of guide RNA “tuning” that enables Cas9 to discriminate betwee...
<p>(A) A schematic diagram showing primers for detecting mutations in Cas9 target site of <i>nsd2</i...
(A) Schematic depicting Cas9 variants and orientation of sgRNAs required to generate blunt, 5’, and ...
<p>a) An in vitro digestion assay shows that sgRNAs differ in their efficiencies. Below the gene nam...
<p>A) Percentage mutation rates (mutant embryos per total number) for maternal Cas9 supply (heterozy...
We report that engineered Cas9 variants with improved specificity-eCas9-1.1 and Cas9-HF1-are often p...
(A) Coding sequence of CASP9 targeted by sgRNA3 used in this work. The CASP9-sgRNA-specific region (...
<p>(A) Schematic showing the recombinant adenoviral genome-harbored <i>EGFP</i> (ADV-EGFP), with the...
The Streptococcus pyogenes Cas9 (SpCas9) nuclease can be efficiently targeted to genomic loci by mea...