Current Issues in Molecular Biology

192 KiB

Open AccessReview

Ethical Issues Regarding CRISPR Mediated Genome Editing

by Zabta Khan Shinwari, Faouzia Tanveer and Ali Talha Khalil

Curr. Issues Mol. Biol. 2018, 26(1), 103-110; https://doi.org/10.21775/cimb.026.103 - 7 Sep 2017

Cited by 47 | Viewed by 13322

CRISPR/Cas9 has emerged as a simple, precise and most rapid genome editing technology. With a number of promising applications ranging from agriculture and environment to clinical therapeutics, it is greatly transforming the field of molecular biology. However, there are certain ethical, moral and [...] Read more.

CRISPR/Cas9 has emerged as a simple, precise and most rapid genome editing technology. With a number of promising applications ranging from agriculture and environment to clinical therapeutics, it is greatly transforming the field of molecular biology. However, there are certain ethical, moral and safety concerns related to the attractive applications of this technique. The most contentious issues concerning human germline modifications are the challenges to human safety and morality such as risk of unforeseen, undesirable effects in clinical applications particularly to correct or prevent genetic diseases, matter of informed consent and the risk of exploitation for eugenics. Stringent regulations and guidelines as well as worldwide debate and awareness are required to ensure responsible and wise use of CRISPR mediated genome editing technology. There is a need for an extensive dialogue among scientists, ethicists, industrialists and policy makers on its societal implications. The opinion of different elements of the society including the general public as well as religious scholars is also critical. In countries with existing legislative framework, it might be appropriate to allow CRISPR-based research to proceed with proper justification. However, much anticipated future clinical applications must be strictly regulated with newly established regulations. Full article

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Open AccessArticle

Applications of CRISPR/Cas9 in Reproductive Biology

by Faheem Ahmed Khan, Nuruliarizki Shinta Pandupuspitasari, Huang ChunJie, Hafiz Ishfaq Ahmad, Kai Wang, Muhammad Jamil Ahmad and ShuJun Zhang

Curr. Issues Mol. Biol. 2018, 26(1), 93-102; https://doi.org/10.21775/cimb.026.093 - 7 Sep 2017

Cited by 5 | Viewed by 1226

Abstract

Genome editing is unravelling its benefits in wide areas of scientific development and understanding. The advances of genome editing from ZFNs and TALLENs to CRISPRs defines its wide applicability. Reproduction is the fundamental process by which all organisms maintain their generations. CRISPR/Cas9, a [...] Read more.

Genome editing is unravelling its benefits in wide areas of scientific development and understanding. The advances of genome editing from ZFNs and TALLENs to CRISPRs defines its wide applicability. Reproduction is the fundamental process by which all organisms maintain their generations. CRISPR/Cas9, a new versatile genome editing tool has been recently tamed to correct several disease causing genetic mutations, spreading its arms to improve reproductive health. It not only edits harmful genetic mutations but is also applied to control the spread of parasitic diseases such as malaria by introducing selfish genetic elements, propagated through generations and population via reproduction. These applications led us to review the recent developments of CRISPRs use in reproductive biology. Full article

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Open AccessReview

CRISPR Mediated Genome Engineering and its Application in Industry

by Saeed Kaboli and Hasan Babazada

Curr. Issues Mol. Biol. 2018, 26(1), 81-92; https://doi.org/10.21775/cimb.026.081 - 7 Sep 2017

Cited by 16 | Viewed by 1286

Abstract

The CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (CRISPR-associated nuclease 9) method has been dramatically changing the field of genome engineering. It is a rapid, highly efficient and versatile tool for precise modification of genome that uses a guide RNA (gRNA) to target [...] Read more.

The CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (CRISPR-associated nuclease 9) method has been dramatically changing the field of genome engineering. It is a rapid, highly efficient and versatile tool for precise modification of genome that uses a guide RNA (gRNA) to target Cas9 to a specific sequence. This novel RNAguided genome-editing technique has become a revolutionary tool in biomedical science and has many innovative applications in different fields. In this review, we briefly introduce the Cas9-mediated genome-editing tool, summarize the recent advances in CRISPR/Cas9 technology to engineer the genomes of a wide variety of organisms, and discuss their applications to treatment of fungal and viral disease. We also discuss advantageous of CRISPR/Cas9 technology to drug design, creation of animal model, and to food, agricultural and energy sciences. Adoption of the CRISPR/Cas9 technology in biomedical and biotechnological researches would create innovative applications of it not only for breeding of strains exhibiting desired traits for specific industrial and medical applications, but also for investigation of genome function. Full article

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Open AccessReview

Improving CRISPR-Cas9 On-Target Specificity

by Muhammad Jamal, Arif Ullah, Muhammad Ahsan, Rohit Tyagi, Zeshan Habib and Khaista Rehman

Curr. Issues Mol. Biol. 2018, 26(1), 65-80; https://doi.org/10.21775/cimb.026.065 - 7 Sep 2017

Cited by 11 | Viewed by 978

Abstract

The CRISPR/Cas9 has revolutionized the field of molecular biology, medical genetics and medicine. The technology is robust, facile and simple to achieve genome targeting in cells and organisms. However, to propagate these nucleases for therapeutic application, the on-target specificity is of paramount importance. [...] Read more.

The CRISPR/Cas9 has revolutionized the field of molecular biology, medical genetics and medicine. The technology is robust, facile and simple to achieve genome targeting in cells and organisms. However, to propagate these nucleases for therapeutic application, the on-target specificity is of paramount importance. Although the binding and cleavage of off-target sites by Cas9 is an issue of concern, however, the specificity of CRISPR technology is greatly improved in current research employing the use of engineer nucleases, improved gRNA selection, novel Cas9 orthologues and the advancement in methods to detect and screen offtarget sites and its effects. Here we summarize the advances in this state-of-the-art technology that will equip the genome editing tools to be applied in clinical research. The researcher should optimize these methods with emphasis to achieving perfection in the specificity. Full article

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Open AccessReview

CRISPR/Cas9-Mediated Immunity in Plants Against Pathogens

by Muhammad Sameeullah, Faheem Ahmed Khan, Göksel Özer, Noreen Aslam, Ekrem Gurel, Mohammad Tahir Waheed and Turan Karadeniz

Curr. Issues Mol. Biol. 2018, 26(1), 55-64; https://doi.org/10.21775/cimb.026.055 - 7 Sep 2017

Cited by 3 | Viewed by 822

Abstract

Global crop production is highly threatened due to pathogen invasion. The huge quantity of pesticides application, although harmful to the environment and human health, is carried out to prevent the crop losses worldwide, every year. Therefore, understanding the molecular mechanisms of pathogenicity and [...] Read more.

Global crop production is highly threatened due to pathogen invasion. The huge quantity of pesticides application, although harmful to the environment and human health, is carried out to prevent the crop losses worldwide, every year. Therefore, understanding the molecular mechanisms of pathogenicity and plant resistance against pathogens is important. The resistance against pathogens is regulated by three important phytohormones, viz. salicylic acid (SA), jasmonic acid ( JA) and ethylene (ET). Here we review the possible role of CRISPR technology to understand the plant pathogenicity by mutating genes responsible for pathogen invasion or up-regulating the phytohormones genes or resistant genes. Thus hormone biosynthesis genes, receptor and feeding genes of pathogens could be important targets for modifications using CRISPR/Cas9 following multiplexing tool box strategy in order to edit multiple genes simultaneously to produce super plants. Here we put forward our idea that the genes would be either mutated in case of plant receptor protein targets of pathogens or up-regulation of resistant genes or hormone biosynthesis genes will be better choice for resistance against pathogens. Full article

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Open AccessReview

An Era of CRISPR/ Cas9 Mediated Plant Genome Editing

by Haris Khurshid, Sohail Ahmad Jan, Zabta Khan Shinwari, Muhammad Jamal and Sabir Hussain Shah

Curr. Issues Mol. Biol. 2018, 26(1), 47-54; https://doi.org/10.21775/cimb.026.047 - 7 Sep 2017

Cited by 9 | Viewed by 942

Abstract

Recently the engineered nucleases have revolutionized genome editing to perturb gene expression at specific sites in complex eukaryotic genomes. Three important classes of these genome editing tools are Zinc Finger Nucleases (ZFN), Meganucleases and Transcription Activator-Like Effector Nucleases (TALEN) which work as hybrid [...] Read more.

Recently the engineered nucleases have revolutionized genome editing to perturb gene expression at specific sites in complex eukaryotic genomes. Three important classes of these genome editing tools are Zinc Finger Nucleases (ZFN), Meganucleases and Transcription Activator-Like Effector Nucleases (TALEN) which work as hybrid systems comprising of target-specific DNA binding domains and molecular scissors or restriction endonucleases. Moreover, the more recent type II clustered regularly inter-spaced short palindromic repeats (CRISPR)-associated protein (CRISPR/ Cas9) system has become the favourite plant genome editing tool for its precision and RNA based specificity unlike its counterparts which rely on protein based specificity. Plasmid-mediated co-delivery of multiple sgRNAs and Cas9 to the plant cell can simultaneously alter more than one target loci which enables multiplex genome editing. In this review, we discuss recent advancements in the CRISPR/Cas9 technology mechanism, theory and its applications in plants and agriculture. We also suggest that the CRISPR/Cas9 as an effective genome editing tool, has vast potential for crop improvement and studying gene regulation mechanisms and chromatin remodelling. Full article

344 KiB

Open AccessReview

Treating Genetic Disorders Using State-Of-The-Art Technology

by Muhammad Jamal, Arif Ullah, Muhammad Ahsan, Rohit Tyagi, Zeshan Habib, Faheem Ahmad Khan and Khaista Rehman

Curr. Issues Mol. Biol. 2018, 26(1), 33-46; https://doi.org/10.21775/cimb.026.033 - 7 Sep 2017

Cited by 1 | Viewed by 708

Abstract

CRISPR/Cas9 [clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9], basically a bacterial immune system, is now widely applicable to engineer genomes of a number of cells and organisms because of its simplicity and robustness. In research avenue the system has been optimized [...] Read more.

CRISPR/Cas9 [clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9], basically a bacterial immune system, is now widely applicable to engineer genomes of a number of cells and organisms because of its simplicity and robustness. In research avenue the system has been optimized to regulate gene expression, modify epigenome and edit target locus. These applications make CRISPR/Cas9 a technology of choice to edit disease causing mutations as well as the epigenome more efficiently than ever before. Meanwhile its application in in vivo and ex vivo cells is encouraging the scientific community for more vigorous gene therapy and in clinical setups for therapeutic genome editing. Here we review the recent advances that CRISPR/Cas9 mediated genome editing has achieved and is reported in previous studies and address the challenges associated with it. Full article

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Open AccessReview

dCas9: A Versatile Tool for Epigenome Editing

by Daan J.W. Brocken, Mariliis Tark-Dame and Remus T. Dame

Curr. Issues Mol. Biol. 2018, 26(1), 15-32; https://doi.org/10.21775/cimb.026.015 - 7 Sep 2017

Cited by 56 | Viewed by 3931

Abstract

The epigenome is a heritable layer of information not encoded in the DNA sequence of the genome, but in chemical modifications of DNA or histones. These chemical modifications, together with transcription factors, operate as spatiotemporal regulators of genome activity. Dissecting epigenome function requires [...] Read more.

The epigenome is a heritable layer of information not encoded in the DNA sequence of the genome, but in chemical modifications of DNA or histones. These chemical modifications, together with transcription factors, operate as spatiotemporal regulators of genome activity. Dissecting epigenome function requires controlled site-specific alteration of epigenetic information. Such control can be obtained using designed DNA-binding platforms associated with effector domains to function as targeted transcription factors or epigenetic modifiers. Here, we review the use of dCas9 as a novel and versatile tool for fundamental studies on epigenetic landscapes, chromatin structure and transcription regulation, and the potential of this approach in basic research in these fields. Full article

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Open AccessReview

Type III CRISPR/Cas System: Introduction and Its Application for Genetic Manipulations

by Tao Liu, Saifu Pan, Yingjun Li, Nan Peng and Qunxin She

Curr. Issues Mol. Biol. 2018, 26(1), 1-14; https://doi.org/10.21775/cimb.026.001 - 7 Sep 2017

Cited by 23 | Viewed by 1932

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) genes provide adaptive immunity against invasion of foreign nucleic acids in archaea and bacteria. The system functions in three distinct stages: adaptation, biogenesis, and interference. CRISPR-Cas systems are currently classified into at least [...] Read more.

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) genes provide adaptive immunity against invasion of foreign nucleic acids in archaea and bacteria. The system functions in three distinct stages: adaptation, biogenesis, and interference. CRISPR-Cas systems are currently classified into at least five different types, each with a signature protein among which Type III systems exhibit a dual DNA/RNA interference activity. Structures of a few Type III surveillance complexes have been determined: they are composed of several different subunits and exhibit striking architectural similarities to Type I surveillance complexes. Here, we review the genetic, biochemical, and structural studies concerning CRISPR-Cas Type III systems and discuss their application for genetic manipulations, including genome engineering and gene silencing. Full article

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Curr. Issues Mol. Biol., Volume 26, Issue 1 (April 2018) – 9 articles

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