key: cord-0926533-nl51y6bz
authors: McCarthy, Matthew W.
title: Harnessing the potential of CRISPR-based platforms to advance the field of hospital medicine
date: 2020-05-04
journal: Expert Rev Anti Infect Ther
DOI: 10.1080/14787210.2020.1761333
sha: 7dd2e6c02841a9825ce5fa560ddd0ba7296cec85
doc_id: 926533
cord_uid: nl51y6bz

Introduction: Clustered regularly interspaced short palindromic repeats (CRISPR) are segments of nucleic acid that play a role in prokaryotic defense and form the basis of a genome editing technology that allows permanent alteration of genetic material. This methodology, known as CRISPR-Cas9, is poised to revolutionize molecular biology, but no literature yet exists on how these advances will affect hospitalists. Areas covered: These specialists in inpatient medicine care for a wide variety of hospitalized patients, including those with infectious disease, cancer, cardiovascular disease, autoimmune disease, hematologic disease, and a variety of other conditions that may soon be impacted by advances in gene-modifying technology provided by CRISPR-Cas9. A Literature search was performed using PubMed [1 December 2019–17 April 2020]. Expert opinion: This paper reviews the remarkable diagnostic and therapeutic potential of the CRISPR-Cas9 platform and concludes with a look at ethical issues and technical hurdles pertaining to the implementation of permanent gene modification in the practice of Hospital Medicine.

Clustered regularly interspaced short palindromic repeats (CRISPR) are segments of nucleic acid containing short, repetitive base sequences that play an important role in prokaryotic defense and form the basis of a genome editing technology that allows permanent modification of genes known as CRISPR-Cas9, as well as CRISPR-Cas12a and CRISPR-13 [1] [2] [3] . While much has been written about how this novel methodology will contribute to scientific inquiry, no literature yet exists on how these advances will affect hospitalistsspecialists in inpatient medicinewho care exclusively for hospitalized patients suffering from a variety of maladies [4] [5] [6] . This paper reviews the potential of the CRISPR platform and explores how this technology may soon impact the field of Hospital Medicine, which has emerged as the fastest growing subspecialty in the history of medicine [7] . It concludes with a look at ethical issues and technical hurdles pertaining to the implementation of permanent gene modification in clinical practice.

The CRISPR-Cas system evolved in microbes as an immune system in prokaryotes that confers resistance to foreign genetic elements such as those present within plasmids and phages [8, 9] . The Cas9 protein performs gene interrogation by unwinding foreign deoxyribonucleic acid (DNA) and determining whether the sequence is complementary to the 20-base pair spacer region of the guide ribonucleic acid (RNA) [10, 11] .

The CRISPR-Cas9 system can be used to manipulate and excise stretches of DNA that have profound relevance for human health, ranging from aggressive malignancies to mitochondrial storage diseases, as well as potential applications in the prevention and treatment of infectious diseases such as human immunodeficiency virus, influenza virus, and malaria, among many others [12] [13] [14] [15] [16] . Other genetic modification systems exist, such as CRISPR-12a and CRISPR-13, but they are outside of the scope of this manuscript, which will focus on CRISPR-Cas9.

Initial applications of gene modification have dealt primarily with conditions that are treated by subspecialty providers (oncologists, rheumatologists, infectious disease specialists), but the CRISPR-Cas9 platform also has profound relevance for primary providers, including pediatricians, obstetricians/ gynecologists, primary care physicians, and hospitalists, which will be the focus of this manuscript.

The term 'hospitalist' was coined in 1996 by Watcher and Goldman and since that time, Hospital Medicine has emerged as the fastest growing subspecialty in the history of medicine [17] . There are currently more than 60,000 hospitalists practicing in the United States, and current professional trends suggest that field will continue its extraordinary growth in the decades ahead [7, 18] . Hospitalists care for a broad array of medical maladies, ranging from blood clots to pneumonia, and emerging research suggests that many of the medical conditions under the hospitalist's purview could be influenced by advances in the CRISPR-Cas9 [19, 20] . Given the rapid evolution of this technology and the widespread appreciation of its potential to alter the treatment of disease, it is necessary for hospitalists to be familiar with the relevance of CRISPR-Cas9. An overview of the most common conditions cared for by hospitalists is provided below followed by an examination of how management of these diseases may change with advances in somatic and germline gene modification [21] .

The emergence of the hospitalist has altered the care of infectious diseases at many hospitals [22] [23] [24] . Patients requiring intravenous antimicrobial therapy are often cared for by hospitalists, with infectious disease specialists serving as consultant (when possible). This means that hospitalists are the primary providers for patients with a wide variety of infectious diseases, which may be broadly divided into the following subgroups: viruses, bacteria, fungi, and parasites [25, 26] . Although these groupings have inherent limitations, they do serve to delineate how CRISPR might be utilized to improve the approach to both diagnosis and treatment.

The CRISPR-Cas9 systems evolved in microbes as a defense mechanism against viruses by cleaving nucleic acids from the invading bacteriophage [27, 28] . For purposes of pathogen detection, cleavage of a labeled sing-strand RNA or DNA probe generates a signal that can provide a fluorescent readout in a number of portable formats, including the use of disposable paper strips, enabling rapid detection of pathogenic viruses [29] . CRISPR-based tests carry several advantages over traditional microbiological diagnostics such as polymerase chain reaction (PCR), which is generally considered to be the most sensitive and specific test available for viral detection. However, PCR require multiple steps to run the assay, including an upfront nucleic acid extraction and amplification steps, and require dedicated instrumentation [30] [31] [32] . By contrast, CRISPR-Cas-based assays can be run directly on primary clinical samples as a single reaction and performed using minimal equipment [33] [34] [35] [36] . This is especially relevant for detection of SARS-CoV-2, where testing has been limited in many places due to a lack of testing materials and personal protective equipment [37] [38] [39] .

As front-line healthcare providers, hospitalists are often the clinicians who must coordinate the work-up of a patient with a suspected infection. Determining the appropriate tests is essential to patient care but also to medical education [40, 41] . Hospitalists have emerged as the primary medical educators on the general medicine service at many academic medical centers and they must be familiar with emerging assays to ensure that patients receive proper treatment and that the next generation of clinicians is properly informed [42, 43] . This includes diagnostic tests for common and emerging viral pathogens.

The SHERLOCK assay has been successfully used to detect both Zika virus and dengue virus directly from bodily fluids, including urine and respiratory samples, in less than two hours and enabled discrimination among viral serotypes [29, 44] . This approach may soon extend to serum samples, and has potential for the rapid diagnosis of human immunodeficiency virus (HIV) as well as hepatitis c virus (HCV) genotype in order to guide the choice of antiviral therapy, which may depend on the presence of single nucleotide polymorphisms [45] [46] [47] [48] .

Others have used the CRISPR-based DETECTR assay to detect human papillomavirus (HPV) types 16 and 18, which are associated with invasive genital tumors, and highlights the potential use of CRISPR-Cas technologies from infectious diseases to other fields, such as cancer (which will be discussed later in this manuscript) [49, 50] . Although more work is necessary to standardize and validate this approach to diagnosis, CRISPR-Cas-based assays show tremendous potential for point-of-care diagnostic because of its ease of implementation, short turnaround time, direct detection from human samples, and colorimetric fluorescent readout [51] [52] [53] .

CRISPR-based platforms also have the potential to detect and alter bacterial pathogens, including those that exist as external threats as well those that live within us and comprise the microbiome [54] . This platform also has tremendous potential to identify and eradicate bacterial resistance genes, which enable pathogens to evade or neutralize antibiotics [2] . One such approach, employed by Quan and colleagues, is FLASH (Finding Low Abundance Sequences by Hybridization), a nextgeneration CRISPR-based diagnostic method that leverages the flexibility and specificity of genetic modification [55] . This approach has been shown to antibiotic resistance genes in saliva and serum and may soon replace multiplex PCR [56] . However, significant hurdles remain. It is unclear how a CRISPR-based platform could remove all clinically-relevant resistance genes in a single bacterium or how this could be scaled up to remove these sequences from an entire population of pathogens.

CRISPR-based methods have also been used to treat drugresistant bacterial infections. In one example, a fifteen-year-old girl with cystic fibrosis with a disseminated Mycobacterium abscessus infection (which included her skin) was treated Article highlights • Clustered regularly interspaced short palindromic repeats (CRISPR) are segments of nucleic acid that play an important role in bacterial defense and form the basis of a genome editing technology known as CRISPR-Cas9. • CRISPR-Cas9 allows for the permanent modification of genetic material and may be harnessed to treat a variety of diseases in humans. • Many of the applications of CRISPR-Cas9 may ultimately affect hospitalists-specialists in inpatient medicine-who serve as the primary caregivers for patients with a wide variety of maladies, ranging from heart disease to cancer. • The CRISPR-Cas9 system has been employed to serve as a diagnostic tool for viral infections. • CRISPR-based human genome editing comes in two forms: heritable germline editing, and non-heritable somatic modifications. with a three-bacteriophage cocktail that had been manipulated using CRISPR [57] . Bacteriophage treatment was associated with marked clinical improvement in this patient who had undergone bilateral lung transplantation and had previously been unable to clear her infection, suggesting that phage therapy may prove to be a useful treatment for drugresistant bacterial infections for other patients, many of whom are now managed by hospitalists at medical centers around the world [12, 58] .

Mycotic infections are broadly divided into those caused by yeasts, molds, and thermally-dimorphic fungi [59] . Many of these organisms are ubiquitous in our environment, and preferentially infect patients with immune impairment [60] . These opportunistic pathogens are on the rise, due to advances in the treatment of cancer and autoimmune conditions, which have produced a variety of new drugs that alter the human immune system [61] . The arsenal of anti-fungal drugs is smallthere are only three major classes of antifungal drugsand novel treatment approaches are desperately needed. One such approach involves CRISPR. Kwon and colleagues provided the first comprehensive analysis and evaluation of different CRISPR approaches for the modification of molds [62] . Guide RNAs were created and CRISPR nucleases were delivered to the filamentous fungus Thermothelomyces thermophilus on plasmids. The team was able to generated high numbers of positive transformants that could be useful for high-throughput assays to identify novel antifungal agents. This approach will likely extend to other fungi, and may yield insights into drug resistance associated with mycotic infection, which have emerged as an important cause of morbidity and mortality in immunocompromised patients.

Parasites encompass a variety of human pathogens [63] . Parasitic worms, such as roundworms and flatworms, are amongst the most complex representing the sixth leading cause of morbidity worldwide and treatment options are limited [64] . Several investigators have employed CRISPR to interrupt essential genes in parasitic worms. For example, in the flatworm Opisthorchis viverrini, CRISPR-induced mutations were introduced into the Ov-grn-1 transcript and protein levels were reduced within 48 h of transfection [65, 66] . The livers of animals infected with modified worms were less swollen, and their biliary ducts had less scarring compared to the those of animals harboring unmodified worms, suggesting that this CRISPR-based method may serve as an approach to attenuate the pathogenicity of a devastating pathogen that affects humans around the world [67, 68] .

The use of CRISPR holds tremendous promise for the treatment of infectious diseases, but many of those assays and therapies are years away. For cancer treatment, CRISPR-based therapies have already arrived [47] . In fact, preliminary results of the first phase-one study using CRISPR to treat cancer were recently revealed. In that study, T-cells from the blood of three patients (two with multiple myeloma and one with sarcoma) and used CRISPR-Cas9 to remove three genes from the cells, including two T-cell receptors as well as programmed cell death-1 (PD-1) [69] . The patients received modified cells after chemotherapy and the CRISPR-edited cells proliferated in all of the patients while no serious treatment-related adverse events were noted. This proof-of-principle study shows that CRISPRbased treatments for cancer are both safe and feasible and will serve as the basis for phase two studies, which will likely begin in the near future [69] [70] [71] [72] . It is not yet known how these malignancies evade immune surveillance or how removal of these genes affected the efficacy of T-cells against cancer.

Another CRISPR-based approach to the treatment of cancer is based on the production of next-generation chimeric antigen receptor (CAR) T-cells, which are created to express cancer-targeting receptors [73, 74] . CAR T-cell therapy targeting the CD19 antigen has been the most studied and successful due to its specific expression in various forms of leukemia. Recently, investigators treated a patient with aggressive lung cancer with T-cells edited by CRISPR [75] . Clinical trials the safety and efficacy of CAR T-cell immunotherapy for leukemia and lymphoma are ongoing and may soon expand to solid organ malignancies [74, 76, 77] .

Heart disease comes in many forms [78, 79] . So too, do the treatment options. Recently, CRISPR has been presented as a potential method to remove monogenic cardiovascular disorders from the offspring and subsequent generations of affected families [80] . Heritable cardiomyopathies, such as dilated cardiomyopathy, hypertrophic cardiomyopathy, and Duchenne muscular dystrophy, vasculopathies such as Marfan's syndrome, and infiltrative diseases such as amyloidosis, serve potential candidates for clinical applications of CRISPR-based germline genome editing techniques [81, 82] . However, technical and ethical hurdles remain. Among these concerns remains the plight of future generations. Permanently editing the germline will affect unborn children who are unable to consent to such a procedure and its use is currently prohibited [83] [84] [85] . But that may soon change.

As the technology advances, CRISPR-based treatments for heart disease may expand beyond heritable cardiomyopathies. It is easy to foresee how gene-editing may be useful for polygenetic syndromes, including hypercholesterolemia, hypertension, and diabetes, among others [80, 86, 87] . For example, one group used CRISPR-Cas9 genome editing to target genes linked to insulin production, SPRY2, and the functional consequences of SPRY2 knockout and overexpression subsequently assessed using glucose uptake and lipid assays [88] . This work implicates a novel mechanism in the development of glucose intolerance and has profound implications for the treatment of diabetes, which is known to be an important risk factor for heart disease. Research in this area is ongoing [82] .

CRISPR-based platforms have been described as the greatest scientific discovery of the twentieth century. However, access to these methodologies has been largely consigned to research laboratories. It is incumbent upon clinicians to familiarize themselves with CRISPR to both understand how it may benefit patients and to help identify future areas of inquiry [89] . As technical and ethical questions regarding the use of CRISPR in humans continue to mount, hospitalists must take a more active role in the discussions to ensure that relevant stakeholders have a say in determining how this technology is implemented. CRISPR provides tremendous potential but also comes with serious pitfalls, and the next decade will be an important period to determine how best to harness this platform to advance the field of hospital medicine.

CRISPR-based platforms have profoundly altered molecular biology and are poised to redefine the practice of medicine [48, [90] [91] [92] . This paper reviews some of the most promising developments in diagnostics and therapeutics that have pertinence to hospitalistsspecialists in inpatient medicinewho care for patients with a wide variety of maladies, ranging from infectious diseases to cancer to heart disease [93, 94] . It is important for hospitalists to understand this emerging new technology both to ensure that it is used properly and to identify areas where it might be further deployed.

This has become especially relevant with the emergence of the novel coronavirus, SARS-CoV-2. In many medical centers, hospitalists have been the primary physicians responsible for the care of patients with this infection. Diagnostic and therapeutic options have been limited, and hospitalists on the front lines have been forced to make treatment decisions with little or no data. This experience has reinforced the crucial role of the hospitalist in the care of patients with novel infections and it further strengthens the argument that hospitalists must be a part of the future research involving emerging pathogens. Frontline clinicians have a better understanding of the questions pertinent to patient care and can help formulate research projects that will address knowledge gaps. One area that has not yet been full explored is the role of CRISPR-Cas9 in the battle against SARS-CoV-2. Permanent nucleic acid modification may provide unique insights into a pandemic that has already infected more than two million people.

But this area of research comes with certain risks and important caveats [95] . CRISPR-based human genome editing comes in two forms: heritable germline editing, and nonheritable somatic modifications [36, 89] . The former results in genetic changes that are passed on to future generations and is currently prohibited to clinical researchers [21, 96] . In 2019, the World Health Organization recommended against any clinical research on human germline editing until all technical and ethical considerations have been properly vetted and this policy is unlikely to change in the foreseeable future [97] [98] [99] .

In fact, some expert now believe that the most relevant hospital applications of CRISPR-Cas9 involve no editing of human DNA. Rather, they see the editing of the microbiota as a far more promising approach, as it would be non-heritable and non-somatic. DNA manipulation may extend to a variety of human pathogens, and may include deletion of viral DNA and cancer DNA to hamper proliferation of malignant cells. As mentioned above, CRISPR-based diagnosis may hasten detection of emerging viral pathogens, such as SARS-CoV-2.

For now, CRISPR-based clinical research will focus on the identification and alteration of infectious pathogens as well as novel treatment options for a variety of diseases ranging from infections to heart disease and cancer [45, [100] [101] [102] [103] .

Recent research indicates that CRISPR has tremendous potential to improve the care of hematologic diseases, such as sickle cell disease and other heritable hemoglobinopathies, as well as autoimmune conditions, such as lupus [35, 47, [103] [104] [105] . But these opportunities must be weighed against potential downstream effects. CRISRP-based platforms have the potential to alter microbiota, which can be associated with adverse health events [106, 107] . It will be crucial for hospitalists, who often care for these patients, to take a more involved role in this aspect of clinical research as CRISPR-based approaches will undoubtedly affect many of their patients in the years to come.

This paper was not funded.

M McCarthy has served as a paid consultant to Allergan. The author has no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Peer reviewers on this manuscript have no relevant financial or other relationships to disclose.

Papers of special note have been highlighted as either of interest (•) or of considerable interest (••) to readers

CRISPR provides acquired resistance against viruses in prokaryotes

An excellent overview of the way CRISPR evolved

Diversity, activity, and evolution of CRISPR loci in Streptococcus thermophilus

Comparative analysis of CRISPR loci in lactic acid bacteria genomes

The impact of hospitalists on medical education and the academic health system

• A superb paper on the ways that hospitalist have affected patient care and medical education

Physician views on caring for hospitalized patients and the hospitalist model of inpatient care

Implications of the hospitalist model for medical students' education

Zero to 50,000 -the 20th anniversary of the hospitalist

The size of the immune repertoire of bacteria

CRISPR-Cas systems can functionally complement each other in DNA defense while providing target flexibility

Harnessing the CRISPR/Cas9 system to disrupt latent HIV-1 provirus

Diversity of CRISPR-Cas immune systems and molecular machines

Current application of CRISPR/Cas9 gene-editing technique to eradication of HIV/AIDS

Use of the CRISPR/Cas9 system as an intracellular defense against HIV-1 infection in human cells

• A look at how CRISPR interacts with nucleic acids

CRISPR/Cas9-mediated CCR5 ablation in human hematopoietic stem/progenitor cells confers HIV-1 resistance in vivo

CRISPR/Cas9 edited NSG mice as PDX models of human leukemia to address the role of niche-derived SPARC

CRISPR-Cas9 gene repair of hematopoietic stem cells from patients with X-linked chronic granulomatous disease

• An important paper on the way that stem cells may be manipulated with CRISPR

The emerging role of "hospitalists" in the American health care system

The hospitalist movement 10 years later: life as a Swiss army knife

CRISPR technologies for stem cell engineering and regenerative medicine

Advances in CRISPR-Cas systems for RNA targeting, tracking and editing

Human germline genome editing

The evolution of the hospitalist model in the United States

•• An excellent overview of hospital medicine in the United States

The hospitalist movement 5 years later

Effects of hospitalist attending physicians on trainee satisfaction with teaching and with internal medicine rotations

Hospitalists as teachers

The positive impact of initiation of hospitalist clinician educators

C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector

RNA targeting with CRISPR-Cas13

Nucleic acid detection with CRISPR-Cas13a/C2c2

Non-invasive methods to diagnose fungal infections in pediatric patients with hematologic disorders

Rapid detection of fungal pathogens in bronchoalveolar lavage samples using panfungal PCR combined with high resolution melting analysis

Evaluation of the Xpert Carba-R (Cepheid) assay using contrived bronchial specimens from patients with suspicion of ventilator-associated pneumonia for the detection of prevalent carbapenemases

Towards a novel therapy against AIDS

ESCRT-II functions by linking to ESCRT-I in HIV-1 budding

Development of gene editing strategies for human β-globin (HBB) gene mutations

• An important look at how gene editing can affect the management of disease in humans

Beyond Mendelian genetics: anticipatory biomedical ethics and policy implications for the use of CRISPR together with gene drive in humans

Era of molecular diagnosis for pathogen identification of unexplained pneumonia, lessons to be learned

Novel 2019 coronavirus SARS-CoV-2 (COVID-19): an updated overview for emergency clinicians

As COVID-19 cases, deaths and fatality rates surge in Italy, underlying causes require investigation

The rise of hospitalists: an opportunity for clinical ethics

Teaching clinical ethics at the bedside: Osler and the essential role of the hospitalist

Frequency of ethical issues on a hospitalist teaching service at an urban, tertiary care center

• An overview of ethical issues facing hospitalists

The rise of hospitalists: an opportunity for infectious diseases investigators

Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6

Nucleic acid detection of plant genes using CRISPR-Cas13

SHERLOCK: nucleic acid detection with CRISPR nucleases

Sharpening the molecular scissors: advances in gene-editing technology. iScience

Quest to use CRISPR against disease gains ground

CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity

In vivo ovarian cancer gene therapy using CRISPR-Cas9

CRISPR tools for systematic studies of RNA regulation

Programmable inhibition and detection of RNA viruses using Cas13

Structural basis for the promiscuous PAM recognition by Corynebacterium diphtheriae Cas9

Challenges and opportunities of nontraditional approaches to treating bacterial infections

FLASH: a next-generation CRISPR diagnostic for multiplexed detection of antimicrobial resistance sequences

•• An intriguing look at the ways that CRISPR could be used in molecular diagnostics

CRISPR-Cas systems exploit viral DNA injection to establish and maintain adaptive immunity

Engineered bacteriophages for treatment of a patient with a disseminated drug-resistant Mycobacterium abscessus

CRISPR editing in biological and biomedical investigation

Future research priorities in fungal resistance

Mold infections of the central nervous system

Novel agents and drug targets to meet the challenges of resistant fungi

Practical guidance for the implementation of the CRISPR genome editing tool in filamentous fungi

Functional genomics tools for haemonchus contortus and lessons from other helminths

Welcome to the CRISPR zoo

Can CRISPR help in the fight against parasitic worms? Elife

Terror in the dirt: sensory determinants of host seeking in soil-transmitted mammalian-parasitic nematodes

The Tao survivorship of schistosomes: implications for schistosomiasis control

Prospects and challenges of CRISPR/Cas genome editing for the study and control of neglected vector-borne nematode diseases

First-in-human phase 1 CRISPR gene editing cancer trials: are we ready?

Emerging role of CRISPR/Cas9 technology for microRNAs editing in cancer research

Genome editing approaches: manipulating of lovastatin and taxol synthesis of filamentous fungi by CRISPR/Cas9 system

CRISPR knock out CTLA-4 enhances the anti-tumor activity of cytotoxic T lymphocytes

CAR-T cells secreting BiTEs circumvent antigen escape without detectable toxicity

Anti-BCMA CAR T-cell therapy bb2121 in relapsed or refractory multiple myeloma

CRISPR gene-editing tested in a person for the first time

•• A landmark paper describing how CRISPR was first tested in a human

CAR-based approaches to cutaneous T-cell lymphoma

Dynamic profiling of antitumor activity of CAR T cells using micropatterned tumor arrays

Syncope: evaluation and differential diagnosis

Global, regional, and national burden of rheumatic heart disease

Genome editing for the understanding and treatment of inherited cardiomyopathies

rs953413 regulates polyunsaturated fatty acid metabolism by modulating ELOVL2 expression. iScience

A patient-centric CRISPR-Cas9 in vitro cardiovascular safety model to examine genetic variability in calcium handling and its impact on drug-induced cardiotoxicity

Crowdsourcing the moral limits of human gene editing?

Ethical issues of CRISPR technology and gene editing through the lens of solidarity

Towards a CRISPR view of early human development: applications, limitations and ethical concerns of genome editing in human embryos

An excellent paper on the ethical issues associated with the use of CRISPR in editing human embryos

Genetic and functional implications of an exonic TRIM55 variant in heart failure

CRISPR-mediated activation of endogenous gene expression in the postnatal heart

CRISPR-Cas9-mediated knockout of SPRY2 in human hepatocytes leads to increased glucose uptake and lipid droplet accumulation

Ethical and scientific issues of gene-edited twin by clustered regularly interspaced short palindromic repeats (CRISPR) Cas9 technology

Genetic predisposition to colon and rectal adenocarcinoma is mediated by a super-enhancer polymorphism coactivating CD9 and PLEKHG6

Ex vivo/in vivo gene editing in hepatocytes using "all-in-one" CRISPR-adeno-associated virus vectors with a self-linearizing repair template. iScience

The Lancet. The better edge of the CRISPR blade

The approved gene therapy drugs worldwide: from 1998 to 2019

Applications of genome editing technology in animal disease modeling and gene therapy

Genome editing: promoting responsible research

What is prudent governance of human genome editing?

The consent form in the Chinese CRISPR study: in search of ethical gene editing

How should physicians respond when they learn patients are using unapproved gene editing interventions?

LEGAL AND ETHICAL IMPLICATIONS OF CRISPR APPLICATIONS IN PSYCHIATRY

Knockout of beta-2 microglobulin reduces stem cell-induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR-24/Bim pathway

A cytosine deaminase for programmable single-base RNA editing

PD-1hiCXCR5-T peripheral helper cells promote B cell responses in lupus via MAF and IL-21

Perspectives of sickle cell disease stakeholders on heritable genome editing

Genetic variations in A20 DUB domain provide a genetic link to citrullination and neutrophil extracellular traps in systemic lupus erythematosus

Beneficial effects of LRP6-CRISPR on prevention of alcohol-related liver injury surpassed fecal microbiota transplant in a rat model

Recording mobile DNA in the gut microbiota using an Escherichia coli CRISPR-Cas spacer acquisition platform