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      • Record: found
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      Cannabis sativa: Interdisciplinary Strategies and Avenues for Medical and Commercial Progression Outside of CBD and THC

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          Abstract

          Cannabis sativa ( Cannabis) is one of the world’s most well-known, yet maligned plant species. However, significant recent research is starting to unveil the potential of Cannabis to produce secondary compounds that may offer a suite of medical benefits, elevating this unique plant species from its illicit narcotic status into a genuine biopharmaceutical. This review summarises the lengthy history of Cannabis and details the molecular pathways that underpin the production of key secondary metabolites that may confer medical efficacy. We also provide an up-to-date summary of the molecular targets and potential of the relatively unknown minor compounds offered by the Cannabis plant. Furthermore, we detail the recent advances in plant science, as well as synthetic biology, and the pharmacology surrounding Cannabis. Given the relative infancy of Cannabis research, we go on to highlight the parallels to previous research conducted in another medically relevant and versatile plant, Papaver somniferum (opium poppy), as an indicator of the possible future direction of Cannabis plant biology. Overall, this review highlights the future directions of cannabis research outside of the medical biology aspects of its well-characterised constituents and explores additional avenues for the potential improvement of the medical potential of the Cannabis plant.

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          A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.

          Martin Jinek, Krzysztof Chylinski, Ines Fonfara … (2012)
          Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. We show here that in a subset of these systems, the mature crRNA that is base-paired to trans-activating crRNA (tracrRNA) forms a two-RNA structure that directs the CRISPR-associated protein Cas9 to introduce double-stranded (ds) breaks in target DNA. At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand, whereas the Cas9 RuvC-like domain cleaves the noncomplementary strand. The dual-tracrRNA:crRNA, when engineered as a single RNA chimera, also directs sequence-specific Cas9 dsDNA cleavage. Our study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.
          Article 0 comments Cited 2964 times – based on 0 reviews     Review now
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            Genome engineering using the CRISPR-Cas9 system.

            F. Ran, Patrick D Hsu, Jason Wright … (2013)
            Targeted nucleases are powerful tools for mediating genome alteration with high precision. The RNA-guided Cas9 nuclease from the microbial clustered regularly interspaced short palindromic repeats (CRISPR) adaptive immune system can be used to facilitate efficient genome engineering in eukaryotic cells by simply specifying a 20-nt targeting sequence within its guide RNA. Here we describe a set of tools for Cas9-mediated genome editing via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, as well as generation of modified cell lines for downstream functional studies. To minimize off-target cleavage, we further describe a double-nicking strategy using the Cas9 nickase mutant with paired guide RNAs. This protocol provides experimentally derived guidelines for the selection of target sites, evaluation of cleavage efficiency and analysis of off-target activity. Beginning with target design, gene modifications can be achieved within as little as 1-2 weeks, and modified clonal cell lines can be derived within 2-3 weeks.
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              RNA-guided human genome engineering via Cas9.

              P. Mali, L. YANG, K. Esvelt … (2013)
              Bacteria and archaea have evolved adaptive immune defenses, termed clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems, that use short RNA to direct degradation of foreign nucleic acids. Here, we engineer the type II bacterial CRISPR system to function with custom guide RNA (gRNA) in human cells. For the endogenous AAVS1 locus, we obtained targeting rates of 10 to 25% in 293T cells, 13 to 8% in K562 cells, and 2 to 4% in induced pluripotent stem cells. We show that this process relies on CRISPR components; is sequence-specific; and, upon simultaneous introduction of multiple gRNAs, can effect multiplex editing of target loci. We also compute a genome-wide resource of ~190 K unique gRNAs targeting ~40.5% of human exons. Our results establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering.
              Article 0 comments Cited 1414 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Pavel B. Drašar: Role: Academic Editor
                Raffaele Capasso: Role: Academic Editor
                Journal
                Journal ID (nlm-ta): Biomedicines
                Journal ID (iso-abbrev): Biomedicines
                Journal ID (publisher-id): biomedicines
                Title: Biomedicines
                Publisher: MDPI
                ISSN (Electronic): 2227-9059
                Publication date (Electronic): 26 February 2021
                Publication date Collection: March 2021
                Volume: 9
                Issue: 3
                Electronic Location Identifier: 234
                Affiliations
                [1 ]Centre for Plant Science, University of Newcastle, University Drive, Callaghan, NSW 2308, Australia; Jackson.Oultram@ 123456uon.edu.au (J.M.J.O.); Joseph.Pegler@ 123456uon.edu.au (J.L.P.); Andy.Eamens@ 123456newcastle.edu.au (A.L.E.)
                [2 ]CannaPacific Pty Ltd., 109 Ocean Street, Dudley, NSW 2290, Australia; tim@ 123456cannapacific.com.au
                [3 ]School of Psychological Sciences, University of Tasmania, Hobart, TAS 7005, Australia; luke.ney@ 123456utas.edu.au
                Author notes
                [* ]Correspondence: Chris.Grof@ 123456newcastle.edu.au ; Tel.: +612-4921-5858
                Author information
                https://orcid.org/0000-0001-7353-3850
                https://orcid.org/0000-0002-2126-8538
                https://orcid.org/0000-0002-6327-3618
                Article
                Publisher ID: biomedicines-09-00234
                DOI: 10.3390/biomedicines9030234
                PMC ID: 7996784
                PubMed ID: 33652704
                SO-VID: 43811b35-df16-48a6-9d70-bf2261c61b84
                Copyright © © 2021 by the authors.
                License:

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 01 February 2021
                Date accepted : 23 February 2021
                Categories
                Subject: Review

                Keywords: cannabis sativa (cannabis),cannabinoids,tetrahydrocannabinol (thc),cannabidiol (cbd),cannabinoid receptors (cb1 and cb2),papaver somniferum (opium poppy),secondary metabolites
                Data availability:
                Keywords: cannabis sativa (cannabis), cannabinoids, tetrahydrocannabinol (thc), cannabidiol (cbd), cannabinoid receptors (cb1 and cb2), papaver somniferum (opium poppy), secondary metabolites

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