geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Rationally engineered Cas9 nucleases with improved specificity


, : Rationally engineered Cas9 nucleases with improved specificity. Science 351(6268): 84-88

The RNA-guided endonuclease Cas9 is a versatile genome-editing tool with a broad range of applications from therapeutics to functional annotation of genes. Cas9 creates double-strand breaks (DSBs) at targeted genomic loci complementary to a short RNA guide. However, Cas9 can cleave off-target sites that are not fully complementary to the guide, which poses a major challenge for genome editing. Here, we use structure-guided protein engineering to improve the specificity of Streptococcus pyogenes Cas9 (SpCas9). Using targeted deep sequencing and unbiased whole-genome off-target analysis to assess Cas9-mediated DNA cleavage in human cells, we demonstrate that "enhanced specificity" SpCas9 (eSpCas9) variants reduce off-target effects and maintain robust on-target cleavage. Thus, eSpCas9 could be broadly useful for genome-editing applications requiring a high level of specificity.

US$19.90

PMID: 26628643

DOI: 10.1126/science.aad5227


Other references

Tan, S.; Lin, Y.; Foo, K.; Koh, H.Fang.; Tow, C.; Zhang, Y.; Ang, L.Wei.; Cui, L.; Badaruddin, H.; Ooi, P.Lim.; Lin, R.Tzer.Pin.; Cutter, J., 2016: Group B Streptococcus Serotype III Sequence Type 283 Bacteremia Associated with Consumption of Raw Fish, Singapore. NlmCategory="UNASSIGNED">We conducted a retrospective study of 40 case-patients and 58 controls as part of a nationwide investigation of a group B Streptococcus outbreak in Singapore in 2015. Eating a Chinese-style raw fish dish (yush...

Hasan, M.A.; Chattopadhyay, P.K., 1997: Varietal difference in the peel pigmentation and quality of litchi (Litchi chinensis Sonn.). Fruit colour is an important quality trait for consumer preference in litchi [Litchi chinensis]. Twelve varieties were evaluated for anthocyanin content, total soluble solids (TSS) and acidity on 8 harvest dates from May to June 1993-94. Bedana an...

Mato, S.; Diaz Cosin, D.J.; Calvin, E.B., 1984: Earthworms in Cabaninos wood (Lumbricidae, Oligochaeta); Sierra de los Ancares, Lugo, Spain. The species composition and abundance of earthworm populations in a woodland area of Lugo province, Spain, are related to soil characteristics and vegetation type. A total of eleven species were recognised in the area. Three facies are identified:...

Vargas, J.Rafael.; Muiser, J., 2014: Promoting universal financial protection: a policy analysis of universal health coverage in Costa Rica (1940-2000). This paper explores the implementation and sustenance of universal health coverage (UHC) in Costa Rica, discussing the development of a social security scheme that covered 5% of the population in 1940, to one that finances and provides comprehensi...

Anisimov, P.I.; Popov, I.A.; Kokushkin, A.M., 2003: The range of pathogen variation due to discussion of different hypotheses of plaque. The paper reviews the hypotheses that explain the mechanism of plague enzooty in natural foci, which are based on a concept of a wide range of plague microbial variability. A comparative analysis of the parameters of variability in the experimenta...

Schrenk, M.O.; Huber, J.A.; Edwards, K.J., 2011: Microbial provinces in the subseafloor. The rocks and sediments of the oceanic subsurface represent a diverse mosaic of environments potentially inhabited by microorganisms. Understanding microbial ecosystems in subseafloor environments confounds standard ecological descriptions in part...

Kruleva, M.; Daskalov, S., 1978: Comparative cytoembryological studies of cytoplasmatic and gene male sterility in the pepper (Capsicum annuum). Doklady 1(8): 1073-1076

Mavrogenis, A.F.; Gambarotti, M.; Angelini, A.; Palmerini, E.; Staals, E.L.; Ruggieri, P.; Papagelopoulos, P.J., 2012: Chondrosarcomas revisited. Chondrosarcomas are malignant bone tumors with pure hyaline cartilage differentiation; myxoid changes, calcification, or ossification may be present. Several subtypes of chondrosarcomas exist. Behavior patterns vary, ranging from slow-growing nonm...

Lincoff A.M.; Popma J.J.; Ellis S.G.; Hacker J.A.; Topol E.J., 1990: Abrupt vessel closure following coronary angioplasty clinical angiographic and therapeutic profile. Circulation 82(4 SUPPL 3): III189

Werner Hans, 1952: Ein neuer Fund der Schnecke Tudicla rusticula in Schleswig-Holstein. Records the second find of the lower Miocene gastropod Tudicla rusticula in Schleswig-Holstein, Germany. Its close affinity with forms of the Bordeaux basin, France, again raises the question of the extent of the lower Miocene sea.