...CRISPR machinery, Quadros et al. recommend ribonucleoprotein (RNP) complexes of Cas9 protein and the separated...alleles using long ssDNA donors and CRISPR ribonucleoproteins.” Genome Biol. 18 (2017) (1):92. DOI: 10.1186...situ germline genome engineering using CRISPR nucleases. Genome Biol. 19 (2018) (1):25. DOI: 10.1186/s13059...Homology Directed Repair Learn about Cas9 Ribonucleoprotein (RNP) Delivery Resources on Addgene.org ...

...came epigenome-engineering tools - zinc finger nucleases and TALENs fused to epigenetic modifiers enabled...Christopher Newgard’s Lab has fused LSD1 to the smaller nuclease-dead Staphylococcus aureus Cas9 (Sa-Cas) and deposited... expression and dCas9-based epigenome editing. Nucleic Acids Research 47:e23–e23. https://doi.org/10.1093...CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Research 44:5615–5628 . https://doi.org/10.1093...

...incorporate multiple single-strand DNA (ssDNA) oligonucleotides (oligos), and thereby rapidly create desired...transient suppression of DNA repair during oligonucleotide integration, allowing MAGE in otherwise unmodified...2016. PubMed PMID: 26884157 Aronshtam, et al. Nucleic Acids Res. 1996. PubMed PMID: 8692687 Tracking... the IL6ST and BACH2 promoters. Vojta A, et al. Nucl. Acids Res. 2016. PubMed PMID: 26969735 Updated...26912792 FusX-TALEN assembly system TALEs as nucleases (TALENs) are genome-editing tools that are deployed...fusion proteins in a target cell. In addition to nucleotide-based linking strategies, post translational ...

...uses the direct injection of a CRISPR/Cas9 ribonucleoprotein complex with linear DNA as repair templates...non-homologous sequence or a series of single-nucleotide substitutions along a 1.5 kb tract, all without...recombination donors in cultured Drosophila cells. Nucleic Acids Research 42:e89–e89 . https://doi.org/10.1093...elegans by CRISPR-targeted homologous recombination. Nucleic Acids Research 41:e193–e193 . https://doi.org/10.1093...

... Cas3. Cas3 functions as both a helicase and a nuclease, unwinding and cutting up DNA after recruitment...consider to be an integral component of CRISPR — a nuclease effector. However, IV-C does in fact have a helicase...Classification and evolution of type II CRISPR-Cas systems. Nucleic Acids Research, 42(10), 6091–6105. https://doi.... and involved in competition between plasmids. Nucleic Acids Research, 48(4), 2000–2012. https://doi.org...

...enhance HDR repair using a single-stranded oligodeoxynucleotide HDR template covalently attached to the ...the Cas9-gRNA RNP complex via the fused HUH endonuclease PCV. This increases HDR efficiency up to 30-fold...meningitidis, Nme2Cas9, that recognizes a simple dinucleotide PAM (N4CC), lending to high target site density...

...experiments of Kim and colleagues using zinc finger nucleases to harness non-homologous end joining repair (...chromosomal deletions in human cells using zinc finger nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. ... interspaced palindromic repeats (CRISPR)/Cas9 nuclease system in mammalian cells. J.Biol.Chem. 2014 Aug...

...activation. Multiple nuclear localization signals (NLS) increase protein import into the nucleus. In developing...-molecule imaging deep inside the cell, in the nucleus and cytoplasm. Again, they found that SunTag marked...

...By mutating one of two Cas9 nuclease domains, researchers created the CRISPR nickase. Nickases create...insert an EcoRI site using a single-stranded oligonucleotide (ssODN) donor. Using a D10A nickase to insert...small insertions or tags (ssODN; IDT Ultramer® Oligonucleotides) Use 100 nt homology arms for large insertions...

...its endonuclease activity to generates a ssDNA substrate which is then nicked by its endonuclease domain...of Rep78 and Rep68 contains a DNA-binding and endonuclease domains, all four Rep proteins share a common...

...genome. Cas genes, which code for polymerases, nucleases, and helicases, were also disrupted to elucidate...CRISPR loci to generate RNA, which guide a Cas nuclease to the specified exogenous genetic material next...mutation in the PAM sequence, preventing the Cas nucleases from re-identifying it (3). Surprisingly, few ...

... Cas9 and a gRNA, which subsequently enter the nucleus and cleave the specified target. Figure adapted...expression is by nature transient, which may lower nuclease toxicity or the potential for off-target effects... Biol. 2000 Jul;10(7):290-5. PubMed. Is VP22 nuclear homing an artifact? Lundberg M, Johansson M. Nat...

...of happen with Cas13. CRISPR-Csm degrades both nuclear and cytoplasmic mRNA with 90-99% knockdown efficiency...CRISPR-Csm presents an attractive alternative for nuclear-RNA targeting, where siRNAs are ineffective and...expression changes of non-target genes and RNA-seq of a nuclear (MALAT1) and cytoplasmic (CKB1) RNA target. Adapted...

...of anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-...anti-FLAG-FB-GFP labeling various targets with nuclear (top, 4x-FLAG-mCherry-H2B), cytoskeletal (middle, 4xFLAG-mCherry-β-actin... editing, while gene editing can be done via a nuclease-active CasMINI. This small yet powerful system...

...and does not consider the missing or damaged nucleotides that are common to biological sources of DSBs...factors, including polymerases (Pol μ and Pol λ), nucleases (Artemis), and structure-specific end cleaning...the Cas9 DSB (blunt or near-blunt ends without nucleotide damage) such products are rare, probably accounting...

...editors with editing windows as small as 1-2 nucleotides! To reduce off-target effects associated with...they purified HF-BE3 protein for delivery in ribonucleoprotein particles (RNPs) to both zebrafish embryos...species. This toolkit includes TALEN and CRISPR nucleases, nickases, and gene activation systems as well... the amount of time that Cas9 is active in the nucleus to reduce off-target gene editing. The bacterial...

... and its organization within the nucleus, also called the 4D nucleome. Visualizing specific loci has historically...loci, a major benchmark in characterizing the 4D nucleome. CRISPRainbow: Scaling up to 6 colors + white ...

... of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299...effect of ammonium sulfate on the activity of ribonuclease A. Biochimica et Biophysica Acta (BBA) - Enzymology...

... As a genome conversion in the transduced cell nucleus, the single-to-double stranded conversion of the...of symmetry. After uncoating in the target cell nucleus, this DNA structure can readily fold into transcriptionally...DNA folds back on itself to become dsDNA in the nucleus and is a good option if you need rapid transgene...

...rarely circulates, but upon viral infection, this nucleic acid can build up. The foreign dsRNA activates ...Transposable elements such as long/short interspersed nuclear elements can generate dsRNA by bidirectional transcription...constantly occurring throughout a single cell, so these nucleic acid hybrids are always around. R-loops are particularly...