Precision Control of CRISPR-Cas9 Using Small Molecules and Light

Biochemistry, Journal Year: 2019, Volume and Issue: 58(4), P. 234 - 244

Published: Jan. 14, 2019

The CRISPR (clustered regularly interspaced short palindromic repeat)-Cas system is an adaptive immune of bacteria that has furnished several RNA-guided DNA endonucleases (e.g., Cas9) are revolutionizing the field genome engineering. Cas9 being used to effect genomic alterations as well in gene drives, where a particular trait may be propagated through targeted species population over generations. ease targeting catalytically impaired any loci led development technologies for base editing, chromatin imaging and modeling, epigenetic regulation. Unsurprisingly, developed numerous applications biotechnology biomedical research therapy agent multiple pathologies. There need precise control activity dimensions, including those dose, time, space these applications. Such precision controls, which required therapeutic agents, particularly important off-target effects, chromosomal translocations, immunogenic response, genotoxicity, embryonic mosaicism observed at elevated levels with prolonged Cas9. Here, we provide perspective on advances aforementioned dimensions using external stimuli small molecules or light) controlled activation, inhibition, degradation

Language: Английский

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DNA methylation in the pathogenesis of polycystic ovary syndrome

Reproduction, Journal Year: 2019, Volume and Issue: 158(1), P. R27 - R40

Published: April 8, 2019

Polycystic ovary syndrome (PCOS) is the leading endocrine and metabolic disorder in premenopausal women characterized by hyperandrogenism abnormal development of ovarian follicles. To date, PCOS etiology remains unclear has been related to insulin resistance, obesity, type 2 diabetes mellitus, cardiovascular disease infertility, among other morbidities. Substantial evidence illustrates impact genetic, intrauterine environmental factors on etiology. Lately, epigenetic have garnered considerable attention pathogenesis considering that changes content DNA methylation, histone acetylation noncoding RNAs reported various tissues with this disease. methylation changed peripheral umbilical cord blood, as well adipose tissue PCOS, suggesting involvement modification Perhaps, these defects promote deregulation genes involved inflammation, hormone synthesis signaling glucose lipid metabolism. Research role just beginning, several issues await investigation. This review aims provide an overview current research focused discuss perspectives regarding topic.

Language: Английский

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Class 2 CRISPR/Cas: an expanding biotechnology toolbox for and beyond genome editing

Cell & Bioscience, Journal Year: 2018, Volume and Issue: 8(1)

Published: Nov. 12, 2018

Artificial nuclease-dependent DNA cleavage systems (zinc-finger nuclease, ZFN; transcription activator like effectors, TALENs) and exogenous nucleic acid defense (CRISPR/Cas) have been used in the new era for genome modification. The most widely toolbox editing, modulation detection contains Types II, V VI of CRISPR/Cas Class 2 systems, categorized characterized by Cas9, Cas12a Cas13 respectively. In this review, we (1) elaborate on definition, classification, structures systems; (2) advance our understanding molecular mechanisms recent progress their applications, especially beyond genome-editing applications; (3) provide insights specificity, efficiency versatility each tool; (4) enhancement specificity toolbox. expanding concerted usage tools is making them more powerful editing other biotechnology applications.

Language: Английский

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TALENs—an indispensable tool in the era of CRISPR: a mini review

Journal of Genetic Engineering and Biotechnology, Journal Year: 2021, Volume and Issue: 19(1), P. 125 - 125

Published: Aug. 21, 2021

Genome of an organism has always fascinated life scientists. With the discovery restriction endonucleases, scientists were able to make targeted manipulations (knockouts) in any gene sequence organism, by technique popularly known as genome engineering. Though there is a range editing tools, but this era dominated CRISPR/Cas9 tool due its ease design and handling. But, when it comes clinical applications, CRISPR not usually preferred. In review, we will elaborate on structural functional role designer nucleases with emphasis TALENs system. We also present unique features limitations CRISPRs which makes better than CRISPRs. robust technology used target specific DNA modifications organism. programmable endonucleases-based manipulating tools such meganucleases (MN), zinc-finger (ZFNs), transcription activator-like effector (TALENs), clustered regularly interspaced short palindromic repeats associated protein (CRISPR/Cas9), research field experienced tremendous acceleration giving rise modern precision specificity. Though, CRISPR-Cas9 platform successfully gained more attention scientific world, ZFNs are their own ways. Apart from high-specificity, proven mitochondrial (mito-TALEN), where gRNA difficult import. This review talks about goals fulfilled drawbacks provides significant insights into pros cons two most popular mini suggests that, novel opportunities therapeutics being highly sensitive toward modifications. article, briefly explore special that indispensable synthetic biology. great perspective providing true guidance researchers working trait improvement via editing.

Language: Английский

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Advances of epigenetic editing

Current Opinion in Chemical Biology, Journal Year: 2020, Volume and Issue: 57, P. 75 - 81

Published: June 29, 2020

Epigenetic editing refers to the locus-specific targeting of epigenetic enzymes rewrite local landscape an endogenous genomic site, often with aim transcriptional reprogramming. Implementing clustered regularly interspaced short palindromic repeat–dCas9 greatly accelerated advancement editing, yielding preclinical therapeutic successes using a variety enzymes. Here, we review current applications these tools in mammals and shed light on biochemical improvements that facilitate versatile applications.

Language: Английский

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