J. Keith Joung

Last updated

J. Keith Joung is an American pathologist and molecular biologist who holds the Robert B. Colvin Endowed Chair in Pathology [1] at Massachusetts General Hospital and is Professor of Pathology at Harvard Medical School. [2] He is a leading figure in the field of genome editing and has pioneered the development of designer nucleases and sensitive off-target detection methods. [3]

Contents

Education

In 1987, Joung graduated from Harvard College with a bachelor's degree in biochemical sciences. [4] He received an M.D. from Harvard Medical School and a Ph.D. in genetics from Harvard University. [5]

Career

Joung is most well known for his work in genome editing and has contributed to the development of designer nucleases through protein engineering and assays for off-target detection. [6] [7] [8] In the mid-2000s, his research was focused on creating zinc finger nuclease tools for biological research and gene therapy. [6] He was the leader and founder of the Zinc Finger Consortium and co-authored a study on Oligomerized Pool Engineering (OPEN), a publicly available strategy for rapidly constructing multi-finger arrays. [9] [10]

More recently, he contributed to the development of TAL effector, TALENs, and the RNA-guided CRISPR/Cas9 system. In addition to demonstrating the use of the CRISPR/Cas9 system in vivo through the zebrafish model, [11] he pioneered the creation of tools such as GUIDE-seq and CIRCLE-seq to detect nuclease off-targets within the genome. [7] [12] In 2016, his group became one of the first to report engineered high-fidelity CRISPR/Cas9 nucleases (HF1) with no detectable off-target effects. [13]

He is one of the scientific co-founders of Editas Medicine, along with Jennifer Doudna, Feng Zhang, George Church, and David Liu. [14] He is also a co-founder of Beam Therapeutics and Verve Therapeutics. [15] [16] He received the Ho-Am Prize in Medicine in 2022 [17] and the American Society of Gene and Cell Therapy Outstanding Achievement Award in 2023, the society's highest honor. [18]

He has an h-index of 85 according to Semantic Scholar. [19]

Related Research Articles

Gene knockouts are a widely used genetic engineering technique that involves the targeted removal or inactivation of a specific gene within an organism's genome. This can be done through a variety of methods, including homologous recombination, CRISPR-Cas9, and TALENs.

<span class="mw-page-title-main">Insertion (genetics)</span> Type of mutation

In genetics, an insertion is the addition of one or more nucleotide base pairs into a DNA sequence. This can often happen in microsatellite regions due to the DNA polymerase slipping. Insertions can be anywhere in size from one base pair incorrectly inserted into a DNA sequence to a section of one chromosome inserted into another. The mechanism of the smallest single base insertion mutations is believed to be through base-pair separation between the template and primer strands followed by non-neighbor base stacking, which can occur locally within the DNA polymerase active site. On a chromosome level, an insertion refers to the insertion of a larger sequence into a chromosome. This can happen due to unequal crossover during meiosis.

<span class="mw-page-title-main">CRISPR</span> Family of DNA sequence found in prokaryotic organisms

CRISPR is a family of DNA sequences found in the genomes of prokaryotic organisms such as bacteria and archaea. These sequences are derived from DNA fragments of bacteriophages that had previously infected the prokaryote. They are used to detect and destroy DNA from similar bacteriophages during subsequent infections. Hence these sequences play a key role in the antiviral defense system of prokaryotes and provide a form of acquired immunity. CRISPR is found in approximately 50% of sequenced bacterial genomes and nearly 90% of sequenced archaea.

Gene editing may refer to:

Zinc Finger Nucleases (ZFNs) are a type of genome editing tool that use engineered proteins to recognize and cut DNA at specific locations. They were one of the first genome editing tools developed and are still used in some applications. However, ZFNs are more complex and expensive than CRISPR/Cas9, which limits their widespread use.

Zinc-finger nucleases (ZFNs) are artificial restriction enzymes generated by fusing a zinc finger DNA-binding domain to a DNA-cleavage domain. Zinc finger domains can be engineered to target specific desired DNA sequences and this enables zinc-finger nucleases to target unique sequences within complex genomes. By taking advantage of endogenous DNA repair machinery, these reagents can be used to precisely alter the genomes of higher organisms. Alongside CRISPR/Cas9 and TALEN, ZFN is a prominent tool in the field of genome editing.

<i>Fok</i>I

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.

Recombinant adeno-associated virus (rAAV) based genome engineering is a genome editing platform centered on the use of recombinant rAAV vectors that enables insertion, deletion or substitution of DNA sequences into the genomes of live mammalian cells. The technique builds on Mario Capecchi and Oliver Smithies' Nobel Prize–winning discovery that homologous recombination (HR), a natural hi-fidelity DNA repair mechanism, can be harnessed to perform precise genome alterations in mice. rAAV mediated genome-editing improves the efficiency of this technique to permit genome engineering in any pre-established and differentiated human cell line, which, in contrast to mouse ES cells, have low rates of HR.

<span class="mw-page-title-main">Transcription activator-like effector nuclease</span>

Transcription activator-like effector nucleases (TALEN) are restriction enzymes that can be engineered to cut specific sequences of DNA. They are made by fusing a TAL effector DNA-binding domain to a DNA cleavage domain. Transcription activator-like effectors (TALEs) can be engineered to bind to practically any desired DNA sequence, so when combined with a nuclease, DNA can be cut at specific locations. The restriction enzymes can be introduced into cells, for use in gene editing or for genome editing in situ, a technique known as genome editing with engineered nucleases. Alongside zinc finger nucleases and CRISPR/Cas9, TALEN is a prominent tool in the field of genome editing.

<span class="mw-page-title-main">Genome editing</span> Type of genetic engineering

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, and the repair of DSBs through homology-directed recombination (HDR) or non-homologous end joining (NHEJ).

<span class="mw-page-title-main">Genetic engineering techniques</span> Methods used to change the DNA of organisms

Genetic engineering techniques allow the modification of animal and plant genomes. Techniques have been devised to insert, delete, and modify DNA at multiple levels, ranging from a specific base pair in a specific gene to entire genes. There are a number of steps that are followed before a genetically modified organism (GMO) is created. Genetic engineers must first choose what gene they wish to insert, modify, or delete. The gene must then be isolated and incorporated, along with other genetic elements, into a suitable vector. This vector is then used to insert the gene into the host genome, creating a transgenic or edited organism.

<span class="mw-page-title-main">Feng Zhang</span> Chinese-American biochemist

Feng Zhang is a Chinese-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. He is most well known for his central role in the development of optogenetics and CRISPR technologies.

<span class="mw-page-title-main">CRISPR interference</span> Genetic perturbation technique

CRISPR interference (CRISPRi) is a genetic perturbation technique that allows for sequence-specific repression of gene expression in prokaryotic and eukaryotic cells. It was first developed by Stanley Qi and colleagues in the laboratories of Wendell Lim, Adam Arkin, Jonathan Weissman, and Jennifer Doudna. Sequence-specific activation of gene expression refers to CRISPR activation (CRISPRa).

<span class="mw-page-title-main">Epigenome editing</span>

Epigenome editing or epigenome engineering is a type of genetic engineering in which the epigenome is modified at specific sites using engineered molecules targeted to those sites. Whereas gene editing involves changing the actual DNA sequence itself, epigenetic editing involves modifying and presenting DNA sequences to proteins and other DNA binding factors that influence DNA function. By "editing” epigenomic features in this manner, researchers can determine the exact biological role of an epigenetic modification at the site in question.

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.

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, creating a double-stranded chromosomal break (DSB) that summons the cell's DNA repair mechanisms 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.

Since antiretroviral therapy requires a lifelong treatment regimen, research to find more permanent cures for HIV infection is currently underway. It is possible to synthesize zinc finger nucleotides with zinc finger components that selectively bind to specific portions of DNA. Conceptually, targeting and editing could focus on host cellular co-receptors for HIV or on proviral HIV DNA.

<span class="mw-page-title-main">CRISPR gene editing</span> Gene editing method

CRISPR gene editing 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 and/or new ones added in vivo.

BLESS, also known as breaks labeling, enrichment on streptavidin and next-generation sequencing, is a method used to detect genome-wide double-strand DNA damage. In contrast to chromatin immunoprecipitation (ChIP)-based methods of identifying DNA double-strand breaks (DSBs) by labeling DNA repair proteins, BLESS utilizes biotinylated DNA linkers to directly label genomic DNA in situ which allows for high-specificity enrichment of samples on streptavidin beads and the subsequent sequencing-based DSB mapping to nucleotide resolution.

GUIDE-Seq is a molecular biology technique that allows for the unbiased in vitro detection of off-target genome editing events in DNA caused by CRISPR/Cas9 as well as other RNA-guided nucleases in living cells. Similar to LAM-PCR, it employs multiple PCRs to amplify regions of interest that contain a specific insert that preferentially integrates into double-stranded breaks. As gene therapy is an emerging field, GUIDE-Seq has gained traction as a cheap method to detect the off-target effects of potential therapeutics without needing whole genome sequencing.

<span class="mw-page-title-main">Kim Jin-soo (biologist)</span> Korean scientist

Kim Jin-Soo is a chemist, biologist, and entrepreneur. He was CEO and CSO, ToolGen, Inc., is a professor in the Department of Chemistry of Seoul National University and director of the Center for Genome Engineering. His research team has developed and improved several types of programmable nucleases, specifically zinc finger nucleases (ZFNs), TAL effector nucleases (TALENs), and RNA-guided engineered nucleases (RGENs). In 2018, he was a Clarivate Analytics Highly Cited Researcher in the cross-field category and in the biology and biochemistry category in 2019.

References

  1. ↑ "Keith Joung, MD, PhD, was recognized as the inaugural incumbent of the Robert B. Colvin Endowed Chair in Patholog".
  2. ↑ "Joung Laboratory - Massachusetts General Hospital, Boston, MA". massgeneral.org. Retrieved 2016-11-19.
  3. ↑ Nair, Prashant (3 May 2016). "QnAs with Jennifer Doudna". Proceedings of the National Academy of Sciences. 113 (18): 4884–4886. Bibcode:2016PNAS..113.4884N. doi: 10.1073/pnas.1605008113 . PMC   4983843 . PMID   27092008. S2CID   8873309.
  4. ↑ "J Keith Joung MD PhD | Advisory Council | ASGCT - American Society of Gene & Cell Therapy | ASGCT - American Society of Gene & Cell Therapy". Archived from the original on 2019-03-06. Retrieved 2019-03-05.
  5. ↑ "Center for Computational and Integrative Biology".
  6. 1 2 Wade, Nicholas (28 December 2009). "In New Way to Edit DNA, Hope for Treating Disease". The New York Times.
  7. 1 2 Tsai, Shengdar Q.; Zheng, Zongli; Nguyen, Nhu T.; Liebers, Matthew; Topkar, Ved V.; Thapar, Vishal; Wyvekens, Nicolas; Khayter, Cyd; Iafrate, A. John; Le, Long P.; Aryee, Martin J.; Joung, J. Keith (February 2015). "GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases". Nature Biotechnology. 33 (2): 187–197. doi:10.1038/nbt.3117. PMC   4320685 . PMID   25513782.
  8. ↑ "CRISPR Researchers Develop Highly Sensitive Method for Identification of Off-Target Effects in Vivo". 12 September 2018.
  9. ↑ "The Zinc Finger Consortium | Consortium Members". zincfingers.org. Retrieved 2016-11-19.
  10. ↑ Maeder, Morgan L.; Thibodeau-Beganny, Stacey; Osiak, Anna; Wright, David A.; Anthony, Reshma M.; Eichtinger, Magdalena; Jiang, Tao; Foley, Jonathan E.; Winfrey, Ronnie J.; Townsend, Jeffrey A.; Unger-Wallace, Erica; Sander, Jeffry D.; Müller-Lerch, Felix; Fu, Fengli; Pearlberg, Joseph; Göbel, Carl; Dassie, Justin P.; Pruett-Miller, Shondra M.; Porteus, Matthew H.; Sgroi, Dennis C.; Iafrate, A. John; Dobbs, Drena; McCray, Paul B.; Cathomen, Toni; Voytas, Daniel F.; Joung, J. Keith (July 2008). "Rapid 'Open-Source' Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification". Molecular Cell. 31 (2): 294–301. doi:10.1016/j.molcel.2008.06.016. PMC   2535758 . PMID   18657511.
  11. ↑ Hwang, Woong Y.; Fu, Yanfang; Reyon, Deepak; Maeder, Morgan L.; Tsai, Shengdar Q.; Sander, Jeffry D.; Peterson, Randall T.; Yeh, J.-R. Joanna; Joung, J. Keith (March 2013). "Efficient genome editing in zebrafish using a CRISPR-Cas system". Nature Biotechnology. 31 (3): 227–229. doi:10.1038/nbt.2501. PMC   3686313 . PMID   23360964.
  12. ↑ Tsai, Shengdar Q.; Nguyen, Nhu T.; Malagon-Lopez, Jose; Topkar, Ved V.; Aryee, Martin J.; Joung, J. Keith (June 2017). "CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR–Cas9 nuclease off-targets". Nature Methods. 14 (6): 607–614. doi:10.1038/nmeth.4278. PMC   5924695 . PMID   28459458.
  13. ↑ Kleinstiver, Benjamin P.; Pattanayak, Vikram; Prew, Michelle S.; Tsai, Shengdar Q.; Nguyen, Nhu T.; Zheng, Zongli; Joung, J. Keith (January 2016). "High-fidelity CRISPR–Cas9 nucleases with no detectable genome-wide off-target effects". Nature. 529 (7587): 490–495. Bibcode:2016Natur.529..490K. doi:10.1038/nature16526. PMC   4851738 . PMID   26735016.
  14. ↑ "Editas Medicine to develop new class of genome editing therapeutics". 25 November 2013.
  15. ↑ "FOUNDERS AND INVENTORS". Beam Therapeutics. Retrieved 14 December 2022.
  16. ↑ "About Us". Verve Therapeutics. Retrieved 5 July 2021.
  17. ↑ 이인준 (31 May 2022). "32회 삼성호암상…오용근 포스텍 교수 등 6인 수상". Newsis (in Korean). Retrieved 3 June 2022.
  18. ↑ https://asgct.org/publications/news/april-2023/am23-awards-announcement.{{cite web}}: Missing or empty |title= (help)
  19. ↑ "J. Joung". Semantic Scholar . Retrieved 14 December 2022.
This page is based on this Wikipedia article
Text is available under the CC BY-SA 4.0 license; additional terms may apply.
Images, videos and audio are available under their respective licenses.