|
Cas9
Cas9 (CRISPR associated protein 9, formerly called Cas5, Csn1, or Csx12) is a 160 dalton (unit), 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 gene editing, 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 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
CRISPR Gene Editing
CRISPR gene editing (; pronounced like "crisper"; an abbreviation for "clustered regularly interspaced short palindromic repeats") is a genetic engineering technique in molecular biology by which the genomes of living organisms may be modified. It is based on a simplified version of the bacterial CRISPR-Cas9 antiviral defense system. By delivering the Cas9 nuclease complexed with a synthetic guide RNA (gRNA) into a cell, the cell's genome can be cut at a desired location, allowing existing genes to be removed or new ones added ''in vivo''. The technique is considered highly significant in biotechnology and medicine as it enables editing genomes ''in vivo'' and is precise, cost-effective, and efficient. It can be used in the creation of new medicines, agriculture, agricultural products, and genetically modified organisms, or as a means of controlling pathogens and pest control, pests. It also offers potential in the treatment of inherited genetic diseases as well as diseases arisi ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
CRISPR
CRISPR (; acronym of clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found in the genomes of prokaryotic organisms such as bacteria and archaea. Each sequence within an individual prokaryotic CRISPR is derived from a DNA fragment of a bacteriophage that had previously infected the prokaryote or one of its ancestors. These sequences are used to detect and destroy DNA from similar bacteriophages during subsequent infections. Hence these sequences play a key role in the antiviral (i.e. anti- phage) defense system of prokaryotes and provide a form of heritable, acquired immunity. CRISPR is found in approximately 50% of sequenced bacterial genomes and nearly 90% of sequenced archaea. Cas9 (or "CRISPR-associated protein 9") is an enzyme that uses CRISPR sequences as a guide to recognize and open up specific strands of DNA that are complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology ... [...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] |
Protospacer Adjacent Motif
A protospacer adjacent motif (PAM) is a 2–6-base pair DNA sequence immediately following the DNA sequence targeted by the Cas9 nuclease in the CRISPR bacterial adaptive immune system. The PAM is a component of the invading virus or plasmid, but is not found in the bacterial host genome and hence is not a component of the bacterial CRISPR locus. Cas9 will not successfully bind to or cleave the target DNA sequence if it is not followed by the PAM sequence. PAM is an essential targeting component which distinguishes bacterial self from non-self DNA, thereby preventing the CRISPR locus from being targeted and destroyed by the CRISPR-associated nuclease. Spacers/protospacers In a bacterial genome, CRISPR loci contain "spacers" (viral DNA inserted into a CRISPR locus) that in type II adaptive immune systems were created from invading viral or plasmid DNA (called "protospacers"). Upon subsequent invasion, a CRISPR-associated nuclease such as Cas9 attaches to a tracrRNA– crRNA comple ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Gene Drive
A gene drive is a natural process and technology of genetic engineering that propagates a particular suite of genes throughout a population by altering the probability that a specific allele will be transmitted to offspring (instead of the Mendelian 50% probability). Gene drives can arise through a variety of mechanisms. They have been proposed to provide an effective means of genetically modifying specific populations and entire species. The technique can employ adding, deleting, disrupting, or modifying genes. Proposed applications include exterminating insects that carry pathogens (notably mosquitoes that transmit malaria, dengue, and zika pathogens), controlling invasive species, or eliminating herbicide or pesticide resistance. As with any potentially powerful technique, gene drives can be misused in a variety of ways or induce unintended consequences. For example, a gene drive intended to affect only a local population might spread across an entire species. Gene driv ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Jennifer Doudna
Jennifer Anne Doudna (; born February 19, 1964) is an American biochemist who has pioneered work in CRISPR gene editing, and made other fundamental contributions in biochemistry and genetics. She received the 2020 Nobel Prize in Chemistry, with Emmanuelle Charpentier, "for the development of a method for genome editing." She is the Li Ka-shing, Li Ka Shing Chancellor, Chancellor's Professor, Chair Professor in the department of chemistry and the department of molecular and cell biology at the University of California, Berkeley. She has been an investigator with the Howard Hughes Medical Institute since 1997. In 2012, Doudna and Emmanuelle Charpentier were the first to propose that CRISPR-Cas9 (enzymes from bacteria that control microbial immunity) could be used for programmable editing of genomes, which has been called one of the most significant discoveries in the history of biology. Since then, Doudna has been a leading figure in what is referred to as the "CRISPR revolution" ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Feng Zhang
Feng Zhang (; born October 22, 1981) is a Chinese-born American biochemist. Zhang currently holds the James and Patricia Poitras Professorship in Neuroscience at the McGovern Institute for Brain Research and in the Departments of Brain and Cognitive Sciences and Biological Engineering at the Massachusetts Institute of Technology. He also has appointments with the Broad Institute of MIT and Harvard (where he is a core member). He is most well known for his role in the development of optogenetics and CRISPR technologies. Early life and education Zhang was born in China in 1981, where both his parents were computer programmers. At age 11, he moved to Iowa with his mother. He attended Theodore Roosevelt High School and Central Academy in Des Moines, graduating in 2000. He earned his B.A. in chemistry and physics in 2004 from Harvard University, where he worked with Xiaowei Zhuang. He then received his PhD in chemical and biological engineering from Stanford University in 2009 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Emmanuelle Charpentier
Emmanuelle Marie Charpentier (; born 11 December 1968) is a French professor and researcher in microbiology, genetics, and biochemistry. As of 2015, she has been a director at the Max Planck Institute for Infection Biology in Berlin. In 2018, she founded an independent research institute, the Max Planck Unit for the Science of Pathogens. In 2020, Charpentier and American biochemist Jennifer Doudna of the University of California, Berkeley, were awarded the Nobel Prize in Chemistry "for the development of a method for genome editing" (through CRISPR gene editing, CRISPR). This was the first science Nobel Prize ever won by two women only. Early life and education Charpentier's paternal grandfather, surnamed Sinanian, was an Armenians, Armenian who escaped to France during the Armenian Genocide and met his wife in Marseille. She was born in 1968 in Juvisy-sur-Orge in France and studied biochemistry, microbiology, and genetics at the Pierre and Marie Curie University (which became ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Virginijus Šikšnys
Virginijus Šikšnys (born 26 January 1956) is a Lithuanian biochemist and a professor at Vilnius University. He is a chief scientist at the Vilnius University Institute of Biotechnology. Biography Šikšnys studied organic chemistry at Vilnius University, receiving his Bachelor's degree in 1978. He then moved to Lomonosov Moscow State University, where he studied enzyme kinetics and received the Candidate of Sciences degree (equivalent to PhD at that time) in 1983. His PhD advisor was Karel Martinek from, at that time, Czechoslovakia. From 1982 till 1993 he worked at the Institute of Applied Enzymology in Vilnius. In 1993 he was a visiting scientist in Robert Huber’s laboratory at the Max-Planck-Institut für Biochemie, Martinsried, Germany. Since 1995 Šikšnys is the chief scientist and head of the Department of Protein-DNA Interactions at the Vilnius University Institute of Biotechnology, since 2006 – professor at Vilnius University and a member of the Lithuania ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Genetic Engineering
Genetic engineering, also called genetic modification or genetic manipulation, is the modification and manipulation of an organism's genes using technology. It is a set of Genetic engineering techniques, technologies used to change the genetic makeup of cells, including the transfer of genes within and across species boundaries to produce improved or novel organisms. New DNA is obtained by either isolating and copying the genetic material of interest using recombinant DNA methods or by Artificial gene synthesis, artificially synthesising the DNA. A Vector (molecular biology), construct is usually created and used to insert this DNA into the host organism. The first recombinant DNA molecule was made by Paul Berg in 1972 by combining DNA from the monkey virus SV40 with the Lambda phage, lambda virus. As well as inserting genes, the process can be used to remove, or "Gene knockout, knock out", genes. The new DNA can either be inserted randomly or Gene targeting, targeted to a spe ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Non-homologous End Joining
Non-homologous end joining (NHEJ) is a pathway that repairs double-strand breaks in DNA. It is called "non-homologous" because the break ends are directly ligated without the need for a homologous template, in contrast to homology directed repair (HDR), which requires a homologous sequence to guide repair. NHEJ is active in both non-dividing and proliferating cells, while HDR is not readily accessible in non-dividing cells. The term "non-homologous end joining" was coined in 1996 by Moore and Haber. NHEJ is typically guided by short homologous DNA sequences called microhomologies. These microhomologies are often present in single-stranded overhangs on the ends of double-strand breaks. When the overhangs are perfectly compatible, NHEJ usually repairs the break accurately. Imprecise repair leading to loss of nucleotides can also occur, but is much more common when the overhangs are not compatible. Inappropriate NHEJ can lead to translocations and telomere fusion, hallmarks of t ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
