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Off-target Genome Editing
Off-target genome editing refers to nonspecific and unintended genetic modifications that can arise through the use of engineered nuclease technologies such as: clustered, regularly interspaced, short palindromic repeats (CRISPR)-Cas9, transcription activator-like effector nucleases ( TALEN), meganucleases, and zinc finger nucleases (ZFN). These tools use different mechanisms to bind a predetermined sequence of DNA (“target”), which they cleave (or "cut"), creating a double-stranded chromosomal break (DSB) that summons the cell's DNA repair mechanisms (non-homologous end joining ( NHEJ) and homologous recombination ( HR)) and leads to site-specific modifications. If these complexes do not bind at the target, often a result of homologous sequences and/or mismatch tolerance, they will cleave off-target DSB and cause non-specific genetic modifications. Specifically, off-target effects consist of unintended point mutations, deletions, insertions inversions, and translocations. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Genome Editing
Genome editing, or genome engineering, or gene editing, is a type of genetic engineering in which DNA is inserted, deleted, modified or replaced in the genome of a living organism. Unlike early genetic engineering techniques that randomly insert genetic material into a host genome, genome editing targets the insertions to site-specific locations. The basic mechanism involved in genetic manipulations through programmable nucleases is the recognition of target genomic loci and binding of effector DNA-binding domain (DBD), double-strand breaks (DSBs) in target DNA by the restriction endonucleases (FokI and CRISPR associated protein, Cas), and the repair of DSBs through homology-directed recombination (HDR) or non-homologous end joining (NHEJ). History Genome editing was pioneered in the 1990s, before the advent of the common current nuclease-based gene-editing platforms, but its use was limited by low efficiencies of editing. Genome editing with engineered nucleases, i.e. all three m ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
ChIP
Chip may refer to: Food * Chip (snack), thinly sliced and deep-fried gastro item ** Potato chips (US) or crisp (UK) * Chips (fried potato strips) (UK) or french fries (US) (common as a takeout side) * Game chips, thin chip/French fries * Chocolate chip Sports and gaming * Chip (association football), a type of football shot * Chip (golf), a type of golf shot * Casino token, often referred to as a chip * Chip, one of the mascots of the University of Colorado at Boulder * Chip, the mascot of Rhode Island FC People and fictional characters * Chip (name), a list of people and fictional characters with either the given name or nickname * Chip (rapper), English hip hop recording artist born Jahmaal Noel Fyffe in 1990 * King Chip (born 1986), stage name of American hip hop rapper Charles Jawanzaa Worth, formerly known as Chip tha Ripper * Chip Douglas, American songwriter, musician, and record producer Douglas Hatlelid (born 1942) * Chip Taylor, stage name of American songwriter Ja ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Flavobacterium Okeanokoites
''Flavobacterium'' is a genus of Gram-negative, nonmotile and motile, rod-shaped bacteria that consists of 130 recognized species. Flavobacteria are found in soil and fresh water in a variety of environments. Several species are known to cause disease in freshwater fish. ''Flavobacterium psychrophilum'' causes the bacterial cold water disease on salmonids and the rainbow trout fry disease on rainbow trout. '' F. columnare'' causes the cotton-wool disease on freshwater fishes. '' F. branchiophilum'' causes the bacterial gill disease on trout. Another member of this genus, '' F. okeanokoites'' is the original source for the type IIs restriction endonuclease ''Fok''I, used in Zinc finger nuclease Zinc-finger nucleases (ZFNs) are artificial restriction enzymes generated by fusing a zinc finger DNA-binding domain to a nuclease, DNA-cleavage domain. Zinc finger domains can be engineered to target specific desired DNA sequences and this enab ...s and TALENs. Species who are a ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
FokI
The restriction endonuclease Fok1, naturally found in ''Flavobacterium okeanokoites'', is a bacterial type IIS restriction endonuclease consisting of an N-terminal DNA-binding domain and a non sequence-specific DNA cleavage domain at the C-terminal. Once the protein is bound to duplex DNA via its DNA-binding domain at the 5'-GGATG-3' recognition site, the DNA cleavage domain is activated and cleaves the DNA at two locations, regardless of the nucleotide sequence at the cut site. The DNA is cut 9 nucleotides downstream of the motif on the forward strand, and 13 nucleotides downstream of the motif on the reverse strand, producing two sticky ends with 4-bp overhangs. Its molecular mass is 65.4 kDa, being composed of 587 amino acids. DNA-binding domain The recognition domain contains three subdomains (D1, D2 and D3) that are evolutionarily related to the DNA-binding domain of the catabolite gene activator protein which contains a helix-turn-helix. DNA-cleavage domain DNA cl ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Dimer (chemistry)
In chemistry, dimerization is the process of joining two identical or similar Molecular entity, molecular entities by Chemical bond, bonds. The resulting bonds can be either strong or weak. Many symmetrical chemical species are described as dimers, even when the monomer is unknown or highly unstable. The term ''homodimer'' is used when the two subunits are identical (e.g. A–A) and ''heterodimer'' when they are not (e.g. A–B). The reverse of dimerization is often called Dissociation (chemistry), dissociation. When two oppositely-charged ions associate into dimers, they are referred to as ''Bjerrum pairs'', after Danish chemist Niels Bjerrum. Noncovalent dimers Anhydrous carboxylic acids form dimers by hydrogen bonding of the acidic hydrogen and the carbonyl oxygen. For example, acetic acid forms a dimer in the gas phase, where the monomer units are held together by hydrogen bonds. Many OH-containing molecules form dimers, e.g. the water dimer. Dimers that form based on w ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Monomer
A monomer ( ; ''mono-'', "one" + '' -mer'', "part") is a molecule that can react together with other monomer molecules to form a larger polymer chain or two- or three-dimensional network in a process called polymerization. Classification Chemistry classifies monomers by type, and two broad classes based on the type of polymer they form. By type: * natural vs synthetic, e.g. glycine vs caprolactam, respectively * polar vs nonpolar, e.g. vinyl acetate vs ethylene, respectively * cyclic vs linear, e.g. ethylene oxide vs ethylene glycol, respectively By type of polymer they form: * those that participate in condensation polymerization * those that participate in addition polymerization Differing stoichiometry causes each class to create its respective form of polymer. : The polymerization of one kind of monomer gives a homopolymer. Many polymers are copolymers, meaning that they are derived from two different monomers. In the case of condensation polymerizations, t ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Indel
Indel (insertion-deletion) is a molecular biology term for an insertion or deletion of bases in the genome of an organism. Indels ≥ 50 bases in length are classified as structural variants. In coding regions of the genome, unless the length of an indel is a multiple of 3, it will produce a frameshift mutation. For example, a common microindel which results in a frameshift causes Bloom syndrome in the Jewish or Japanese population. Indels can be contrasted with a point mutation. An indel inserts or deletes nucleotides from a sequence, while a point mutation is a form of substitution that ''replaces'' one of the nucleotides without changing the overall number in the DNA. Indels can also be contrasted with Tandem Base Mutations (TBM), which may result from fundamentally different mechanisms. A TBM is defined as a substitution at adjacent nucleotides (primarily substitutions at two adjacent nucleotides, but substitutions at three adjacent nucleotides have been observed). In ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Cas9 Nickases
Cas9 (CRISPR associated protein 9, formerly called Cas5, Csn1, or Csx12) is a 160 kilodalton protein which plays a vital role in the immunological defense of certain bacteria against DNA viruses and plasmids, and is heavily utilized in genetic engineering applications. Its main function is to cut DNA and thereby alter a cell's genome. The CRISPR-Cas9 genome editing technique was a significant contributor to the Nobel Prize in Chemistry in 2020 being awarded to Emmanuelle Charpentier and Jennifer Doudna. More technically, Cas9 is a RNA-guided DNA endonuclease enzyme associated with the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) adaptive immune system in ''Streptococcus pyogenes''. ''S. pyogenes'' utilizes CRISPR to memorize and Cas9 to later interrogate and cleave foreign DNA, such as invading bacteriophage DNA or plasmid DNA. Cas9 performs this interrogation by unwinding foreign DNA and checking for sites complementary to the 20 nucleotide spacer region ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Guide RNA
Guide RNA (gRNA) or single guide RNA (sgRNA) is a short sequence of RNA that functions as a guide for the Cas9-endonuclease or other Cas-proteins that cut the double-stranded DNA and thereby can be used for gene editing. In bacteria and archaea, gRNAs are a part of the CRISPR-Cas system that serves as an adaptive immune defense that protects the organism from viruses. Here the short gRNAs serve as detectors of foreign DNA and direct the Cas-enzymes that degrades the foreign nucleic acid. History The RNA editing guide RNA was discovered in 1990 by B. Blum, N. Bakalara, and L. Simpson through Northern Blot Hybridization in the mitochondrial maxicircle DNA of the eukaryotic parasite Leishmania tarentolae. Subsequent research throughout the mid-2000s and the following years explored the structure and function of gRNA and the CRISPR-Cas system. A significant breakthrough occurred in 2012 when it was discovered that gRNA could guide the Cas9 endonuclease to introduce target-specific cu ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Streptococcus Pyogenes
''Streptococcus pyogenes'' is a species of Gram-positive, aerotolerant bacteria in the genus '' Streptococcus''. These bacteria are extracellular, and made up of non-motile and non-sporing cocci (round cells) that tend to link in chains. They are clinically important for humans, as they are an infrequent, but usually pathogenic, part of the skin microbiota that can cause group A streptococcal infection. ''S. pyogenes'' is the predominant species harboring the Lancefield group A antigen, and is often called group A ''Streptococcus'' (GAS). However, both '' Streptococcus dysgalactiae'' and the '' Streptococcus anginosus'' group can possess group A antigen as well. Group A streptococci, when grown on blood agar, typically produce small (2–3 mm) zones of beta-hemolysis, a complete destruction of red blood cells. The name group A (beta-hemolytic) ''Streptococcus'' is thus also used. The species name is derived from Greek words meaning 'a chain' () of berries ( atiniz ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
