...widely used genome editing tool. However, Cas9’s endonuclease activity can be limited by the need for a specific...and follow the DNA sequence targeted by the Cas9 nuclease. The Streptococcus pyogenes Cas9 (SpCas9) requires... 2-D linear molecule snaking through our cells’ nuclei--our chromatin is comprised of thousands of loops...provide a 3-D structure for packing genomic DNA into nuclei, but are also thought to play a key role in the...

...particles enter the nucleus through nuclear pores and thus do not require breakdown of the nuclear envelope during...during cell division to enter the nucleus as gamma-retroviruses do. Fun fact: Since lentivirus and gamma-retrovirus...

...through nuclear localization signals which can traffic their viral particles inside the nucleus, without...access to the host genome requires breakdown of the nuclear envelope, which occurs during mitosis. The most...PMCID: PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human immunodeficiency virus type 1 preintegration...

... the miCube, SQUID (Hongquan, et. al, 2020) or openUC2 (Diederich et. al., 2020) aim to make microscopy.... There are a number of new platforms (such as openUC2, which I am a co-founder of), that aim to support...microscope or open-source modular optical toolbox openUC2 are trying to overcome this hurdle by taking these...He is co-founder of the open-hardware startup “openUC2” and focuses on bringing cutting-edge research ...

...identifies plasmid features that differ by only a few nucleotides, such as the fluorescent proteins EGFP and mEGFP...listing commonly used primers detected in a given nucleotide sequence. The table includes primer names, sequences...determine similarity between a given sequence and nucleotide (BLASTN) or protein (BLASTX) sequence databases...databases. Additionally, align a custom nucleotide sequence against a given sequence using BLAST2. See this...

...Cpf1 CRISPR Nucleases   Cas9 Cpf1 Components crRNAtracrRNACas9 crRNACpf1 Endonuclease size spCas9...and doesn’t require the expression of other endonucleases, like Csy4, or the inclusion of processing signals...Zhang, F. (2015). Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell,163(3),...

...a CRISPR repeat array near a putative effector nuclease. The Cas13d proteins they identified have little...identified Cas13a-c orthologs, but they do include HEPN nuclease domains characteristic of the Cas13 superfamily...great results with CasRx and AdmCas13d fused to a nuclear localization signal (NLS). In a HEK293 mCherry ...each of four targets, including two mRNAs and two nuclear long non-coding RNAs. One interesting twist to ...

... works best for genome editing  with an active nuclease.  Read the paper in Molecular Cell Find the CasMINI...Programmed genome editing by a miniature CRISPR-Cas12f nuclease Quanjiang Ji’s lab describes an effective miniature...various delivery methods, including plasmid, ribonucleoprotein, and AAV.  Read the paper in Nature Chemical...protect the RNA from degradation by cellular exonucleases and broadly improve editing efficiency. Read...

...a few advantages over other cloning methods. Exonuclease-based methods like Gibson assembly require 20...multiple DNA fragments to create TAL effector nucleases. These plasmids were designed to utilize the BsaI...Gate cloning for inserting the appropriate oligonucleotides specifying a gRNA target sequence into a Cas9... Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic Acids Res. 2011 Jul;39(12):e82. Epub 2011 Apr 14...

... Then, pCRISPR is introduced along with an oligonucleotide repair template. Through recombineering, the...VA, Marraffini LA (2014) Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials. Nat...expression using an engineered CRISPR-Cas system. Nucleic Acids Research 41:7429–7437 . https://doi.org/10.1093...antimicrobials using efficiently delivered RNA-guided nucleases. Nat Biotechnol 32:1141–1145 . https://doi.org...tuberculosisusing an efficient CRISPR interference system. Nucleic Acids Res 44:e143–e143 . https://doi.org/10.1093...

...non-coding RNAs (lncRNA). lncRNAs are over 200 nucleotides in length and lack an open reading frame, so ...activation (CRISPRa) are powerful tools. CRISPRi uses a nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily...Charpentier E. A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012...with the CRISPR/Cas9 system in human cell lines. Nucleic Acids Research. 2014;43(3):e17. doi:10.1093/nar...

...MLV)-like particles loaded with Cas9-gRNA ribonucleoproteins (RNPs). According to the authors, they named...insertion of a DNA sequence ranging from a single nucleotide change to large insertions like the addition ...intracellular injections into the cytoplasm or pronucleus of the zygote which is both technically challenging...viral-derived Nanoblades loaded with Cas9-sgRNA ribonucleoproteins." Nature communications 10.1 (2019): 45. ...

...post-transcriptional additional of multiple adenine (A) nucleotides to the tail of a messenger RNA transcript. The...protection from degradation, and is integral to the nuclear export and translation processes as well.   ...employ other non-poly(A) transcript-stabilizing and nuclear export elements such as WPRE (5) and CTE (6), and... Gag expression in a context-dependent manner. Nucleic Acids Research. 2000. PubMed PMID:10648781. PubMed...

...CRISPR plasmids containing a nuclease into three functional groups: nucleases, modulators, and chimeric ...chimeric effectors. Nucleases generate a classic double-stranded break, while modulators regulate transcription...function, from 2014 to 2023. As you can see, nucleases, the original CRISPR tool, remain the most popular...

...such a system is likely to introduce additional nucleotides flanking the insertion. If the DNA ends anneal...MMEJ may also introduce, substitute, or delete nucleotides in addition to the expected deletion, but the...compare to CRISPR sticky-end insertion using the nuclease Cpf1. Since Cpf1 cuts in a staggered pattern, ...this editing platform lies in the diversity of nucleases and their applications. It will be interesting...

...improve nuclear localization of fluorescent proteins, they found that adding a triple nuclear localization...protease allows a transcription factor to enter the nucleus and modulate transcription of the mCitrine gene...includes 95 plasmids consisting of CRISPR/Cas nucleases, base editors, gRNA backbones, and promoters for...

...fibroblasts and keratinocytes to peripheral blood mononuclear cells (PBMCs). PBMCs isolated from whole blood...behave as separate extrachromosomal elements in the nucleus of the target cell. The method also reduces biosafety...Episomal vectors are based on the Epstein-Barr Nuclear Antigen-1 (oriP-EBNA-1) system where each vector... peripheral blood mononuclear cells...

...further improved, by adding a nuclear localization sequence (SV40 Large T nuclear localization sequence) and...sites. FRT sites and loxP sites differ at the nucleotide level but share an overall structure of 13 base...the fusions allowing Cre/FLP translocate to the nucleus where they exert their recombinase activity (Logie...

...target sequence was digested by sgRNA guided Cas9 endonuclease, homology dependent repair (HR, homologous recombination...however, if the microinjection was done as the pronuclei were fusing, you may have several mosaic mice....off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases." Genome research 24.1 (2014): ...Daisuke, et al. "Generation of mutant mice by pronuclear injection of circular plasmid expressing Cas9...

...can be caused by intracellular factors such as nucleases and reactive oxygen species, or external forces...: The 5’ DNA end of the break is resected by nucleases to create a ssDNA 3’ overhang. This will serve...ssDNA templates (referred to as ssODNs (oligodeoxynucleotides)) are commonly used for smaller modifications...alleles using long ssDNA donors and CRISPR ribonucleoproteins. Quadros RM, et al. Genome Biol. 18(1); (...