Prime Editing
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Prime Editing
Prime editing is a 'search-and-replace' genome editing technology in molecular biology by which the genome of living organisms may be modified. The technology directly writes new genetic information into a targeted DNA site. It uses a fusion protein, consisting of a catalytically impaired Cas9 endonuclease fused to an engineered reverse transcriptase enzyme, and a prime editing guide RNA (pegRNA), capable of identifying the target site and providing the new genetic information to replace the target DNA nucleotides. It mediates targeted Insertion (genetics), insertions, Deletion (genetics), deletions, and base-to-base conversions without the need for double strand breaks (DSBs) or donor DNA templates. The technology is an early-stage, experimental genome editing method that has received mainstream press attention due to its potential uses in medical genetics. It utilizes methodologies similar to precursor genome editing technologies, including CRISPR/Cas9-mediated genome editing, CRIS ...
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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 inserts 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 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, however, its use was limited by low efficiencies of editing. Genome editing with engineered nucleases, i.e. all three major classes of ...
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Transfection
Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. It may also refer to other methods and cell types, although other terms are often preferred: "transformation" is typically used to describe non-viral DNA transfer in bacteria and non-animal eukaryotic cells, including plant cells. In animal cells, transfection is the preferred term as transformation is also used to refer to progression to a cancerous state (carcinogenesis) in these cells. Transduction is often used to describe virus-mediated gene transfer into eukaryotic cells. The word ''transfection'' is a portmanteau of ''trans-'' and ''infection''. Genetic material (such as supercoiled plasmid DNA or siRNA constructs), may be transfected. Transfection of animal cells typically involves opening transient pores or "holes" in the cell membrane to allow the uptake of material. Transfection can be carried out using calcium phosphate (i.e. tricalcium phosphate), by ele ...
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Homology Directed Repair
Homology-directed repair (HDR) is a mechanism in cells to repair double-strand DNA lesions. The most common form of HDR is homologous recombination. The HDR mechanism can only be used by the cell when there is a homologous piece of DNA present in the nucleus, mostly in G2 and S phase of the cell cycle. Other examples of homology-directed repair include single-strand annealing and breakage-induced replication. When the homologous DNA is absent, another process called non-homologous end joining (NHEJ) takes place instead. Cancer suppression HDR is important for suppressing the formation of cancer. HDR maintains genomic stability by repairing broken DNA strands; it is assumed to be error free because of the use of a template. When a double strand DNA lesion is repaired by NHEJ there is no validating DNA template present so it may result in a novel DNA strand formation with loss of information. A different nucleotide sequence in the DNA strand results in a different protein exp ...
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Non-homologous End Joining
Non-homologous end joining (NHEJ) is a pathway that repairs double-strand breaks in DNA. NHEJ is referred to as "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 ...
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CRISPR/Cas9
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 dual 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 ...
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Faculty Opinions
F1000 (formerly "Faculty of 1000") is an open research publisher for scientists, scholars, and clinical researchers. F1000 offers a different research evaluation service from standard academic journals by offering peer-review after, rather than before, publishing a research article. Initially, F1000 was named after the 1,000 faculty members that performed peer-reviews, but over time F1000 expanded to more than 8,000 members. When F1000 was acquired by Taylor & Francis Group in January 2020, it kept the publishing services. F1000Prime (AKA Faculty Opinions) and F1000 Workspace (AKA Sciwheel) were acquired by different brands. History ''Faculty of 1000'' was founded in 2000 by publishing entrepreneur Vitek Tracz in London. Initially, it was named after the 1,000 experts it had reviewing academic works, but over time F1000 expanded to more than 8,000 members. In 2002, it introduced F1000Prime (later known as Faculty Opinions), which recommended scientific articles selected by its ...
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Annual Reviews (publisher)
Annual Reviews is an independent, non-profit academic publishing company based in San Mateo, California. As of 2021, it publishes 51 journals of review articles and ''Knowable Magazine'', covering the fields of life, biomedical, physical, and social sciences. Review articles are usually “peer-invited” solicited submissions, often planned one to two years in advance, which go through a peer-review process. The organizational structure has three levels: a volunteer board of directors, editorial committees of experts for each journal, and paid employees. Annual Reviews' stated mission is to synthesize and integrate knowledge "for the progress of science and the benefit of society". The first Annual Reviews journal, the ''Annual Review of Biochemistry'', was published in 1932 under the editorship of Stanford University chemist J. Murray Luck, who wanted to create a resource that provided critical reviews on contemporary research. The second journal was added in 1939. By ...
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Annual Review Of Genomics And Human Genetics
The ''Annual Review of Genomics and Human Genetics'' is a peer-reviewed scientific journal published by Annual Reviews (publisher), Annual Reviews since 2000. It releases an annual volume of review articles relevant to the fields of genomics and human genetics. Aravinda Chakravarti and Eric D. Green have been the journal's co-editor-in-chief, editors since 2005. As of 2022, it has a 2021 impact factor of 9.340. History The ''Annual Review of Genomics and Human Genetics'' was first published in 2000. The nonprofit publisher Annual Reviews (publisher), Annual Reviews decided that its existing journals of the ''Annual Review of Medicine'' and ''Annual Review of Genetics'' were shifting their coverage from biochemical genetics to molecular interpretation; the new journal could focus on the intersection of genetics and medicine. Another impetus for forming the journal was the then-ongoing Human Genome Project to map the human genome. The founding editor-in-chief, editor was Eric Lander, ...
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Broad Institute
The Eli and Edythe L. Broad Institute of MIT and Harvard (IPA: , pronunciation respelling: ), often referred to as the Broad Institute, is a biomedical and genomic research center located in Cambridge, Massachusetts, Cambridge, Massachusetts, United States. The institute is independently governed and supported as a 501(c)(3) nonprofit research organization under the name Broad Institute Inc., and it partners with the Massachusetts Institute of Technology, Harvard University, and the five Harvard teaching hospitals. History The Broad Institute evolved from a decade of research collaborations among MIT and Harvard scientists. One cornerstone was the Center for Genome Research of Whitehead Institute at MIT. Founded in 1982, the Whitehead became a major center for genomics and the Human Genome Project. As early as 1995, scientists at the Whitehead started pilot projects in genomic medicine, forming an unofficial collaborative network among young scientists interested in genomic a ...
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David R
David (; , "beloved one") (traditional spelling), , ''Dāwūd''; grc-koi, Δαυΐδ, Dauíd; la, Davidus, David; gez , ዳዊት, ''Dawit''; xcl, Դաւիթ, ''Dawitʿ''; cu, Давíдъ, ''Davidŭ''; possibly meaning "beloved one". was, according to the Hebrew Bible, the third king of the United Kingdom of Israel. In the Books of Samuel, he is described as a young shepherd and harpist who gains fame by slaying Goliath, a champion of the Philistines, in southern Canaan. David becomes a favourite of Saul, the first king of Israel; he also forges a notably close friendship with Jonathan, a son of Saul. However, under the paranoia that David is seeking to usurp the throne, Saul attempts to kill David, forcing the latter to go into hiding and effectively operate as a fugitive for several years. After Saul and Jonathan are both killed in battle against the Philistines, a 30-year-old David is anointed king over all of Israel and Judah. Following his rise to power, David ...
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Indels
Indel is a molecular biology term for an insertion or deletion of bases in the genome of an organism. It is classified among small genetic variations, measuring from 1 to 10 000 base pairs in length, including insertion and deletion events that may be separated by many years, and may not be related to each other in any way. A microindel is defined as an indel that results in a net change of 1 to 50 nucleotides. 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 f ...
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