The cytochrome P450 3A (CYP3A) gene family’s role in early progression of gastric cancer was comprehensively investigated. Its potential as a therapeutic target was evaluated. Upon literature review, aberrant expression of the CYP3A gene family has a strong correlation with gastric cancer onset, although the precise underlying mechanisms remain unclear. To assess its potential as a biomarker for early diagnosis and a therapeutic target, we have provided a comprehensive review of the regulatory mechanisms governing CYP3A gene family expression in gastric cancer, as well as its relation with early tumor progression and the tumor microenvironment. The CYP3A gene family is crucial in the proliferation, migration, and invasion of gastric cancer cells and promotes cancer progression by modulating inflammatory responses and oxidative stress within the tumor microenvironment. Furthermore, genetic polymorphisms in CYP3Aenzymes highlight its potential value in personalized medicine. Based on these findings, this paper explores the feasibility of developing inhibitors and activators targeting CYP3A enzymes and discusses potential applications in gene therapy. This research provides crucial theoretical support for the CYP3A gene family as an early diagnostic marker and therapeutic target for gastric cancer. In the future, multi-omics studies and large-scale clinical trials will be essential to advance clinical translation of these findings.

• Gastric cancer
• Cytochrome P450 3A
• Early diagnosis
• Therapeutic target
• Precision medicine

• Blood disorders
• CRISPR-Cas
• Clinical trials
• Delivery methods
• Gene therapy

• Animals
• Humans
• CRISPR-Cas Systems
• Gene Editing/methods
• Genetic Therapy/methods
• Hematologic Diseases/therapy
• Hematologic Diseases/genetics

• CRISPR
• blood coagulation disorders
• gene editing
• haemophilia
• lexicon

• Humans
• Gene Editing/methods
• Hemophilia A/genetics
• Hemophilia A/therapy

• Regeneron Pharmaceuticals, Inc.
• Intellia Therapeutics, Inc

• CRISPR/Cas9
• Exosomes
• Gene therapy

• Exosomes/genetics
• Exosomes/metabolism
• Gene Editing/methods
• CRISPR-Cas Systems
• Humans
• Animals
• Gene Transfer Techniques
• Genetic Therapy/methods

• AAV
• CRISPR-Cas-9
• bispecific antibodies
• gene therapy
• hemophilia
• immune tolerance induction
• viral vectors

• Humans
• Genetic Therapy/methods
• Hemophilia A/therapy
• Hemophilia A/genetics
• Factor VIII/genetics
• Factor VIII/therapeutic use
• Factor IX/genetics
• Hemophilia B/therapy
• Hemophilia B/genetics
• Animals
• Genetic Vectors/genetics

• none Academic Leadership Program Priority 2030 by the Federal State Autonomous Educational Institution of Higher Education I.M. Sechenov, First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University)
• none The grant of the state program of the «Sirius» Federal Territory «Scientific and technological development of the «Sirius» Federal Territory»

• ASXL1
• CircRNA
• cancer

• Humans
• RNA, Circular/genetics
• Epigenesis, Genetic
• Neoplasms/genetics
• Mutation
• Transcription Factors/genetics
• Biomarkers
• Repressor Proteins/genetics

• CRISPR
• Cancer
• Gene editing
• Gene therapy

• Humans
• Gene Editing/methods
• CRISPR-Cas Systems/genetics
• Genetic Therapy/methods

• CRISPR/Cas
• blood disorder
• non-viral vectors
• physical delivery
• viral vectors

Chronic myeloid leukaemia (CML) is a haematological neoplasm driven by the BCR/ABL fusion oncogene. The monogenic aspect of the disease and the feasibility of ex vivo therapies in haematological disorders make CML an excellent candidate for gene therapy strategies. The ability to abolish any coding sequence by CRISPR-Cas9 nucleases offers a powerful therapeutic opportunity to CML patients. However, a definitive cure can only be achieved when only CRISPR-edited cells are selected. A gene-trapping approach combined with CRISPR technology would be an ideal approach to ensure this. Here, we developed a CRISPR-Trap strategy that efficiently inserts a donor gene trap (SA-CMV-Venus) cassette into the BCR/ABL-specific fusion point in the CML K562 human cell line. The trapping cassette interrupts the oncogene coding sequence and expresses a reporter gene that enables the selection of edited cells. Quantitative mRNA expression analyses showed significantly higher level of expression of the BCR/Venus allele coupled with a drastically lower level of BCR/ABL expression in Venus+ cell fractions. Functional in vitro experiments showed cell proliferation arrest and apoptosis in selected Venus+ cells. Finally, xenograft experiments with the selected Venus+ cells showed a large reduction in tumour growth, thereby demonstrating a therapeutic benefit in vivo. This study represents proof of concept for the therapeutic potential of a CRISPR-Trap system as a novel strategy for gene elimination in haematological neoplasms.

• BCR/ABL
• CRISPR
• CRISPR-Trap
• chronic myeloid leukaemia
• gene therapy

• Apoptosis/genetics
• CRISPR-Cas Systems/genetics
• Cell Proliferation/genetics
• Chronic Disease
• Fusion Proteins, bcr-abl/genetics
• Fusion Proteins, bcr-abl/metabolism
• Humans
• Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy
• Leukemia, Myelogenous, Chronic, BCR-ABL Positive/therapy

• CRISPR
• Cas9
• cancer
• gene editing
• guide RNA

• CRISPR-Cas system
• Gene editing
• Gene therapy
• Genome editing
• Meganucleases
• TALEN
• ZFN

• CRISPR-Cas system
• Gene editing
• Gene therapy
• Genome editing
• Meganucleases
• TALEN
• ZFN

Thalassemia is a genetic haematological disorder that arises due to defects in the α and β-globin genes. Worldwide, 0.3-0.4 million children are born with haemoglobinopathies per year. Thalassemic patients, as well as their families, face various serious clinical, socio-economic, and psychosocial challenges throughout their life. Different therapies are available in clinical practice to minimize the suffering of thalassemic patients to some extent and potentially cure the disease. Predominantly, patients undergo transfusion therapy to maintain their haemoglobin levels. Due to multiple transfusions, the iron levels in their bodies are elevated. Iron overload results in damage to body organs, resulting in heart failure, liver function failure or endocrine failure, all of which are commonly observed. Certain drugs have been developed to enhance the expression of the γ-gene, which ultimately results in augmentation of fetal haemoglobin (HbF) levels and total haemoglobin levels in the body. However, its effectiveness is dependent on the genetic makeup of the individual patient. At present, allogeneic haematopoietic Stem Cell Transplantation (HSCT) is the only practically available option with a high curative rate. However, the outcome of HSCT is strongly influenced by factors such as age at transplantation, irregular iron chelation history before transplantation, histocompatibility, and source of stem cells. Gene therapy using the lentiglobin vector is the most recent method for cure without any mortality, graft rejection and clonal dominance issues. However, delayed platelet engraftment is being reported in some patients. Genome editing is a novel approach which may be used to treat patients with thalassemia; it makes use of targeted nucleases to correct the mutations in specific DNA sequences and modify the sequence to the normal wild-type sequence. To edit the genome at the required sites, CRISPR/Cas9 is an efficient and accurate tool that is used in various genetic engineering programs. Genome editing mediated by CRISPR/Cas9 has the ability to restore the normal β-globin function with minimal side effects. Using CRISPR/Cas9, expression of BCL11A can be downregulated along with increased production of HbF. However, these genome editing tools are still under in-vitro trials. CRISPR/Cas9 has can be used for precise transcriptional regulation, genome modification and epigenetic editing. Additional research is required in this regard, as CRISPR/Cas9 may potentially exhibit off-target activity and there are legal and ethical considerations regarding its use.

• CRISPR/Cas9
• base editors
• gene therapy
• genome editing
• haematopoietic stem cell transplant
• thalassemia

• CRISPR/Cas9
• Genome editing
• dermatology
• gene therapy
• microbes
• site mutation.

• CRISPR-Cas9
• Gene editing
• In-vitro
• Research laboratory

Genome editing reemerged in 2012 with the development of CRISPR/Cas9 technology, which is a genetic manipulation tool derived from the defense system of certain bacteria against viruses and plasmids. This method is easy to apply and has been used in a wide variety of experimental models, including cell lines, laboratory animals, plants, and even in human clinical trials. The CRISPR/Cas9 system consists of directing the Cas9 nuclease to create a site-directed double-strand DNA break using a small RNA molecule as a guide. A process that allows a permanent modification of the genomic target sequence can repair the damage caused to DNA. In the present study, the basic principles of the CRISPR/Cas9 system are reviewed, as well as the strategies and modifications of the enzyme Cas9 to eliminate the off-target cuts, and the different applications of CRISPR/Cas9 as a system for visualization and gene expression activation or suppression. In addition, the review emphasizes on the potential application of this system in the treatment of different diseases, such as pulmonary, gastrointestinal, hematologic, immune system, viral, autoimmune and inflammatory diseases, and cancer.

• Animals
• CRISPR-Cas Systems
• DNA Repeat Expansion
• Disease Models, Animal
• Gene Editing/methods
• Gene Editing/trends
• Gene Silencing
• Gene Transfer Techniques
• Genetic Diseases, Inborn/genetics
• Genetic Diseases, Inborn/pathology
• Genetic Diseases, Inborn/therapy
• Humans
• MEDLINE
• Mice
• Neuromuscular Diseases/genetics
• Neuromuscular Diseases/pathology
• Neuromuscular Diseases/therapy

• AAV
• Alpha1 Antitrypsin deficiency
• CRISPR
• Clinical trials
• DMD
• DNA damage
• HDR
• Myotonic Dystrophy
• NHEJ
• nanoparticles

• Animals
• CRISPR-Cas Systems/genetics
• Gene Editing/methods
• Genetic Therapy/methods
• Humans
• Muscular Dystrophy, Duchenne/genetics
• Myotonic Dystrophy/genetics

• R01 EB023776 NIBIB NIH HHS
• R01 HL139605 NHLBI NIH HHS

Advances in Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated system (CRISPR/Cas9) has dramatically reshaped our ability to edit genomes. The scientific community is using CRISPR/Cas9 for various biotechnological and medical purposes. One of its most important uses is developing potential therapeutic strategies against diseases. CRISPR/Cas9 based approaches have been increasingly applied to the treatment of human diseases like cancer, genetic, immunological and neurological disorders and viral diseases. These strategies using CRISPR/Cas9 are not only therapy oriented but can also be used for disease modeling as well, which in turn can lead to the improved understanding of mechanisms of various infectious and genetic diseases. In addition, CRISPR/Cas9 system can also be used as programmable antibiotics to kill the bacteria sequence specifically and therefore can bypass multidrug resistance. Furthermore, CRISPR/Cas9 based gene drive may also hold the potential to limit the spread of vector borne diseases. This bacterial and archaeal adaptive immune system might be a therapeutic answer to previous incurable diseases, of course rigorous testing is required to corroborate these claims. In this review, we provide an insight about the recent developments using CRISPR/Cas9 against various diseases with respect to disease modeling and treatment, and what future perspectives should be noted while using this technology.

• CRISPR/Cas9
• Disease modeling
• Genetic diseases

CRISPR/Cas9 technology was used to abrogate p210 oncoprotein expression in the Boff-p210 cell line, a pro-B line derived from interlukin-3-dependent Baf/3, that shows IL-3-independence arising from the constitutive expression of BCR-ABL p210. Using this approach, pools of Boff-p210-edited cells and single edited cell-derived clones were obtained and functionally studied in vitro. The loss of p210 expression in Boff-p210 cells resulted in the loss of ability to grow in the absence of IL-3, as the Baf/3 parental line, showing significantly increased apoptosis levels. Notably, in a single edited cell-derived clone carrying a frame-shift mutation that prevents p210 oncoprotein expression, the effects were even more drastic, resulting in cell death. These edited cells were injected subcutaneously in immunosuppressed mice and tumor growth was followed for three weeks. BCR/ABL-edited cells developed smaller tumors than those originating from unedited Boff-p210 parental cells. Interestingly, the single edited cell-derived clone was unable to develop tumors, similar to what is observed with the parental Baf/3 cell line.

CRISPR/Cas9 genomic editing technology allows the ablation of the BCR/ABL fusion gene, causing an absence of oncoprotein expression, and blocking its tumorigenic effects in vitro and in the in vivo xenograft model of CML. The future application of this approach in in vivo models of CML will allow us to more accurately assess the value of CRISPR/Cas9 technology as a new therapeutic tool that overcomes resistance to the usual treatments for CML patients.

• BCR/ABL
• CRISPR/Cas9
• genome edition
• leukemia

The revolutionary technology for genome editing known as the clustered regularly interspaced short palindromic repeat (CRISPR)‐CRISPR‐associated protein 9 (Cas9) system has sparked advancements in biological and biomedical research. The scientific breakthrough of the development of CRISPR‐Cas9 technology has allowed us to recapitulate human diseases by generating animal models of interest ranging from zebrafish to non‐human primates. The CRISPR‐Cas9 system can also be used to delineate the mechanisms underlying the development of human disorders and to precisely correct disease‐causing mutations. Repurposing this technology enables wider applications in transcriptome and epigenome manipulation and holds promise to reach the clinic. In this review, we highlight the latest advances of the CRISPR‐Cas9 system in different platforms and discuss the hurdles and challenges this technology is facing. J. Cell. Biochem. 118: 4152–4162, 2017. © 2017 Wiley Periodicals, Inc.

• CRISPR-Cas9
• CRISPRa
• CRISPRi
• DISEASE MODELS
• GENE CORRECTION
• GENOME EDITING
• IMAGING