Gene Editing Technology Research Articles

CRISPR-Cas9 in hiPSCs: A new era in personalized treatment for Stargardt disease

CRISPR-Cas9 in hiPSCs: A new era in personalized treatment for Stargardt disease

Strategies for Reducing Toxicity and Enhancing Efficacy of Chimeric Antigen Receptor T Cell Therapy in Hematological Malignancies.

Chimeric antigen receptor T cell (CAR-T) therapy in hematologic malignancies has made great progress, but there are still some problems. First, T cells from tumor patients show an exhaustion phenotype; thus, the persistence and function of the CAR-Ts are poor, and achieving a satisfactory curative effect is difficult. Second, some patients initially respond well but quickly develop antigen-negative tumor recurrence. Thirdly, CAR-T treatment is not effective in some patients and is accompanied by severe side effects, such as cytokine release syndrome (CRS) and neurotoxicity. The solution to these problems is to reduce the toxicity and enhance the efficacy of CAR-T therapy. In this paper, we describe various strategies for reducing the toxicity and enhancing the efficacy of CAR-T therapy in hematological malignancies. In the first section, strategies for modifying CAR-Ts using gene-editing technologies or combining them with other anti-tumor drugs to enhance the efficacy of CAR-T therapy are introduced. The second section describes some methods in which the design and construction of CAR-Ts differ from the conventional process. The aim of these methods is to enhance the anti-tumor activity of CAR-Ts and prevent tumor recurrence. The third section describes modifying the CAR structure or installing safety switches to radically reduce CAR-T toxicity or regulating inflammatory cytokines to control the symptoms of CAR-T-associated toxicity. Together, the knowledge summarized herein will aid in designing better-suited and safer CAR-T treatment strategies.

Cytosolic Fructose-1,6-bisphosphate Aldolases Modulate Primary Metabolism and Phytohormone Homeostasis in Soybean

Fructose-1,6-bisphosphate aldolase (FBA) is an important catalytic enzyme in carbon metabolism and plays an important role in plant growth and development. Currently, the biological functions of FBA in soybean (Glycine max) remain unknown. In this study, we conducted research on FBA in soybean and identified 14 GmFBA genes. Among them, GmFBAc1 and GmFBAc2 are broadly expressed in different tissues. Double mutant lines of GmFBAc1 and GmFBAc2 were obtained by CRISPR-Cas9 gene editing technology. Compared with the wild type, the double-gene homozygous mutant gmfbac1gmfbac2 exhibited dwarf seedlings and narrow leaflets, indicating that GmFBAc1 and GmFBAc2 are critical for soybean growth and development. The gmfbac1gmfbac2 metabolomic analysis revealed that compared to the wild type, carbohydrate metabolism was reduced and amino acid metabolism was enhanced in gmfbac1gmfbac2 mutant leaves. Transcriptomic analysis showed that genes in IAA signaling and JA signaling were downregulated and upregulated, respectively. Our study demonstrates an important role of GmFBAc1 and GmFBAc2 in modulating carbon metabolism and phytohormone homeostasis.

Modeling Duchenne Muscular Dystrophy Cardiomyopathy with Patients' Induced Pluripotent Stem-Cell-Derived Cardiomyocytes.

Duchenne muscular dystrophy (DMD) is an X-linked progressive muscle degenerative disease caused by mutations in the dystrophin gene, resulting in death by the end of the third decade of life at the latest. A key aspect of the DMD clinical phenotype is dilated cardiomyopathy, affecting virtually all patients by the end of the second decade of life. Furthermore, despite respiratory complications still being the leading cause of death, with advancements in medical care in recent years, cardiac involvement has become an increasing cause of mortality. Over the years, extensive research has been conducted using different DMD animal models, including the mdx mouse. While these models present certain important similarities to human DMD patients, they also have some differences which pose a challenge to researchers. The development of somatic cell reprograming technology has enabled generation of human induced pluripotent stem cells (hiPSCs) which can be differentiated into different cell types. This technology provides a potentially endless pool of human cells for research. Furthermore, hiPSCs can be generated from patients, thus providing patient-specific cells and enabling research tailored to different mutations. DMD cardiac involvement has been shown in animal models to include changes in gene expression of different proteins, abnormal cellular Ca2+ handling, and other aberrations. To gain a better understanding of the disease mechanisms, it is imperative to validate these findings in human cells. Furthermore, with the recent advancements in gene-editing technology, hiPSCs provide a valuable platform for research and development of new therapies including the possibility of regenerative medicine. In this article, we review the DMD cardiac-related research performed so far using human hiPSCs-derived cardiomyocytes (hiPSC-CMs) carrying DMD mutations.

Pathological modeling of glycogen storage disease type III with CRISPR/Cas9 edited human pluripotent stem cells.

Introduction: Glycogen storage disease type III (GSDIII) is a rare genetic disease caused by mutations in the AGL gene encoding the glycogen debranching enzyme (GDE). The deficiency of this enzyme, involved in cytosolic glycogen degradation, leads to pathological glycogen accumulation in liver, skeletal muscles and heart. Although the disease manifests with hypoglycemia and liver metabolism impairment, the progressive myopathy is the major disease burden in adult GSDIII patients, without any curative treatment currently available. Methods: Here, we combined the self-renewal and differentiation capabilities of human induced pluripotent stem cells (hiPSCs) with cutting edge CRISPR/Cas9 gene editing technology to establish a stable AGL knockout cell line and to explore glycogen metabolism in GSDIII. Results: Following skeletal muscle cells differentiation of the edited and control hiPSC lines, our study reports that the insertion of a frameshift mutation in AGL gene results in the loss of GDE expression and persistent glycogen accumulation under glucose starvation conditions. Phenotypically, we demonstrated that the edited skeletal muscle cells faithfully recapitulate the phenotype of differentiated skeletal muscle cells of hiPSCs derived from a GSDIII patient. We also demonstrated that treatment with recombinant AAV vectors expressing the human GDE cleared the accumulated glycogen. Discussion: This study describes the first skeletal muscle cell model of GSDIII derived from hiPSCs and establishes a platform to study the mechanisms that contribute to muscle impairments in GSDIII and to assess the therapeutic potential of pharmacological inducers of glycogen degradation or gene therapy approaches.

Identification of two novel rice S genes through combination of association and transcription analyses with gene-editing technology.

Traditional rice blast resistance breeding largely depends on utilizing typical resistance (R) genes. However, the lack of durable R genes has prompted rice breeders to find new resistance resources. Susceptibility (S) genes are potential new targets for resistance genetic engineering using genome-editing technologies, but identifying them is still challenging. Here, through the integration of genome-wide association study (GWAS) and transcriptional analysis, we identified two genes, RNG1 and RNG3, whose polymorphisms in 3'-untranslated regions (3'-UTR) affected their expression variations. These polymorphisms could serve as molecular markers to identify rice blast-resistant accessions. Editing the 3'-UTRs using CRISPR/Cas9 technology affected the expression levels of two genes, which were positively associated with rice blast susceptibility. Knocking out either RNG1 or RNG3 in rice enhanced the rice blast and bacterial blight resistance, without impacting critical agronomic traits. RNG1 and RNG3 have two major genotypes in diverse rice germplasms. The frequency of the resistance genotype of these two genes significantly increased from landrace rice to modern cultivars. The obvious selective sweep flanking RNG3 suggested it has been artificially selected in modern rice breeding. These results provide new targets for S gene identification and open avenues for developing novel rice blast-resistant materials.

Gene-editing technologies for developing climate resilient rice crops in sub-Saharan Africa: Political priorities and space for responsible innovation

Over the last 2 decades, rice has become one of the most important staple crops for sub-Saharan Africa. Estimates show that average consumption of rice has tripled over the last 3 decades, from 9.2 million metric tons (Mt) in the early 1990s to 31.5 million Mt in 2018, with West and Central Africa accounting for nearly two-thirds of this share. The demand for rice, however, has placed an enormous economic burden on African countries, whereby they spent over USD 5.5 billion per year on rice imports over the past few years. To address this challenge, over 32 countries have established National Rice Development Strategies to increase local production and to achieve rice self-sufficiency. Several of these countries have shown policy interest to use modern biotechnological advancements, including gene editing, to ensure increases in rice productivity and reduce food imports, in the context of extreme climate vulnerability and acceleration of the effects of biotic and abiotic stresses. This review article examines the role of biotechnology in African countries’ efforts to achieve rice self-sufficiency, particularly the potential for genome-editing technologies toward the genetic improvement of rice and to Africa’s nascent research programs. This article notes that while gene editing offers important advances in crop breeding, like genetic engineering, it faces some persistent sociopolitical challenges and low societal acceptability. As such, international partnerships advancing genome editing in Africa’s rice-subsectors development could benefit from adopting key principles from “responsible research and innovation” to help these projects achieve their potential, while bringing about more inclusive and reflexive processes that strive to anticipate the benefits and limits associated with new biotechnologies as they relate to local contexts. Such an approach could create the necessary political space to test and assess the benefits (and risks) related to adopting gene-editing technologies in Africa’s rice sectors.

A CRISPR interference strategy for gene expression silencing in multiple myeloma cell lines

BackgroundMultiple myeloma (MM) is the second most common hematologic neoplasm which is characterized by proliferation and infiltration of plasmatic cells in the bone marrow. Currently, MM is considered incurable due to resistance to treatment. The CRISPR/Cas9 system has emerged as a powerful tool for understanding the role of different genetic alterations in the pathogenesis of hematologic malignancies in both cell lines and mouse models. Despite current advances of gene editing tools, the use of CRISPR/Cas9 technology for gene editing of MM have not so far been extended. In this work, we want to repress Rnd3 expression, an atypical Rho GTPase involved in several cellular processes, in MM cell lines using a CRISPR interference strategy.ResultsWe have designed different guide RNAs and cloning them into a lentiviral plasmid, which contains all the machinery necessary for developing the CRISPR interference strategy. We co-transfected the HEK 293T cells with this lentiviral plasmid and 3rd generation lentiviral envelope and packaging plasmids to produce lentiviral particles. The lentiviral particles were used to transduce two different multiple myeloma cell lines, RPMI 8226 and JJN3, and downregulate Rnd3 expression. Additionally, the impact of Rnd3 expression absence was analyzed by a transcriptomic analysis consisting of 3’ UTR RNA sequencing. The Rnd3 knock-down cells showed a different transcriptomic profile in comparison to control cells.ConclusionsWe have developed a CRISPR interference strategy to generate stable Rnd3 knockdown MM cell lines by lentiviral transduction. We have evaluated this strategy in two MM cell lines, and we have demonstrated that Rnd3 silencing works both at transcriptional and protein level. Therefore, we propose CRISPR interference strategy as an alternative tool to silence gene expression in MM cell lines. Furthermore, Rnd3 silencing produces changes in the cellular transcriptomic profile.

Autophagy and Its Lineage-Specific Roles in the Hematopoietic System.

Autophagy is a dynamic process that regulates the selective and nonselective degradation of cytoplasmic components, such as damaged organelles and protein aggregates inside lysosomes to maintain tissue homeostasis. Different types of autophagy including macroautophagy, microautophagy, and chaperon-mediated autophagy (CMA) have been implicated in a variety of pathological conditions, such as cancer, aging, neurodegeneration, and developmental disorders. Furthermore, the molecular mechanism and biological functions of autophagy have been extensively studied in vertebrate hematopoiesis and human blood malignancies. In recent years, the hematopoietic lineage-specific roles of different autophagy-related (ATG) genes have gained more attention. The evolution of gene-editing technology and the easy access nature of hematopoietic stem cells (HSCs), hematopoietic progenitors, and precursor cells have facilitated the autophagy research to better understand how ATG genes function in the hematopoietic system. Taking advantage of the gene-editing platform, this review has summarized the roles of different ATGs at the hematopoietic cell level, their dysregulation, and pathological consequences throughout hematopoiesis.

Abstract 254: Apolipoprotein B Mediates Hepatic Proliferation And Cell Survival Signalling: A Crispr/cas-9 Gene Editing ApoB-knockout Study

Introduction: Apolipoprotein B (ApoB) is a key structure protein in very-low-density lipoprotein (VLDL). Ample research has been focusing on its crucial role in lipoprotein metabolism and the onset of cardiovascular disease (CVD). However, in vivo research on the broader pathophysiological function of ApoB has been hindered by the embryonic lethality of ApoB knockout (ApoB-KO) animal models. Thus, the objective of this study is to establish ApoB-KO cell models and demonstrate ApoB as a key determinant in governing hepatocyte proliferation and apoptosis. Approaches and Results: the Crispr/Cas-9 gene editing technology was applied to genetically disrupt ApoB expression in both human and mouse hepatocytes. Total RNAs of the parental and ApoB-KO cells were subjected to RNA sequencing profiling for whole transcriptome. Subsequent bioinformatics analysis revealed that ApoB-KO significantly altered expression of more than 2000 genes in hepatocytes. More importantly, expression of genes involved in DNA replication, cell cycle, cell proliferation and apoptotic signaling were the genes mostly affected, which were either up- or down-regulated by more than 5-folds. These included the histone associated genes H2ac10-12, cyclin Ccna2, Ccnb1 and Cdc20, cell proliferation gene Fosb and apoptosis gene P53. A BrDU cell proliferation assay detecting the actively proliferating cells further demonstrated that depletion of ApoB greatly inhabited cell proliferation, indicated by the markedly reduced cell numbers and the prolonged cell doubling time in ApoB-KO cells compared to their parental cells. Mechanistic study revealed that cAMP signaling, and the insulin associated PI3K kinase pathway that medicated gene transcription, cell growth and proliferation were profoundly impaired upon ApoB depletion. Reconstitution of ApoB in the KO cells by expressing a truncated ApoB protein, ApoB72, was able to promote cell growth rate and improve cell survival to a level comparable to the parental cells. Conclusion: This is the first study that characterizes a novel function of ApoB as a key determinant in hepatic cell proliferation and survival pathways. This novel finding greatly advances our knowledge of ApoB beyond the boundary of VLDL metabolism and CVD.

Muller morphs redefined by a multimorphic IRF4 variant presenting as CID with PCP susceptibility

In their last review, the IUIS reported 40 inborn errors of immunity (IEI) leading to combined immunodeficiency (CID). Yet, the genetic cause of CID in a sizeable percentage of patients remains unknown. We describe seven patients with CID from four continents with similar clinical presentations characterized by severe viral and recurrent respiratory infections, notably by Pneumocystis jirovecii. Two of the patients also suffered from mycobacterial disease. All these seven patients carry the heterozygous mutation T95R in the DNA binding domain of the interferon regulatory factor 4 (IRF4). Leveraging state-of-the-art transcriptomic, molecular, computational, and gene editing technologies, we discovered that IRF4-T95R does not fit into any single category of the Muller morphs but in three of them simultaneously. In fact, we showed that IRF4-T95R causes a unique combination of (i) gain-of-function by binding DNA with higher affinity than IRF4-WT; (ii) loss-of-function since IRF4-T95R fails to induce canonical IRF4 regulated genes, and; (iii) neomorphic functions by inducing the transcription of genes that IRF4-WT does not. These alterations in the transcriptional programming caused by the mutation lead to cellular and humoral immunological abnormalities. Patients’ B cells displayed impaired maturation, defective plasma cell differentiation, and isotype switching, all which resulted in agammaglobulinemia. Their T cells, despite proliferating normally, showed a decreased production of pro-inflammatory cytokines and a reduced capacity to differentiate into TH17 and TFH. Two different knock-in mouse models recapitulated the patients’ phenotypes. Overall, our data show that autosomal dominant IRF4-T95R is a new genetic cause of CID and defines a novel multimorphic molecular mechanism of a disease characterized by gain, loss, and new functions of IRF4.

Archaeal and bacteriophage derived Cas proteins demonstrate memory T cell response in humans

Abstract The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas system is a powerful gene editing tool with clinical applications in the pipeline. In vivodelivery of Cas proteins could induce immune responses which would limit the utility of the technology. Previously, we and others have demonstrated pre-existing immunity to Cas9 proteins derived from Staphylococcus aureus(Sa) and Streptococcus pyogenes(Sp) in human populations. As these Cas proteins are of microbial origin this finding was not surprising. There are numerous naturally occurring alternatives to Cas9 and many of these are being leveraged for gene editing. Alternatives to Cas9 that have successfully been used in gene editing technologies include Cas12a (derived from Acidaminococcus sp), Cas14a (derived from DPANN, a super-phylum of archaea) and Casϕ (derived from a huge bacteriophage). It has been postulated that humans would not have pre-existing immunity to some of these Cas proteins. In the comparative study described here, we evaluated pre-existing antibodies and memory T-cell responses to Cas9, Cas12a, Cas14a and Casϕ from the same 18 healthy donors for all proteins. Pre-existing antibodies were identified using ELISA and memory T-cell responses were measured using the ELISpot assay. All four Cas proteins tested positive for both assays in at least some donors. It has been speculated that pre-existing immunity to Cas9 proteins is due to human exposure to the pathogenic bacteria they are derived from. Consequently, Cas14a and Casϕ should not be associated with pre-existing immunity. However, our results show that anti-Cas14a and anti-Casϕ antibodies were detected in ~50% of the study population and these donors also showed memory T-cell responses.

Development and Prospect of Gene Editing Technology

Gene editing technology, as an important technology in the field of science, can accurately and stably modify the genome sequence, bringing revolutionary changes to various fields of life sciences, so it has attracted wide attention from researchers in various countries. This article makes a brief introduction to ZFN and TALEN technology, and at the same time makes a detailed introduction to CRISPR/Cas9 technology, focusing on the limitations, application, and development of CRISPR/Cas9 system. This research found that the CRISPR/Cas9 system and its related technologies are going deep into various fields of life sciences and have achieved remarkable results, providing more ideas and possibilities for human disease treatment, crop gene control and other fields.

The Impact of CRISPR and Cas9 on AIDS

AIDS is a chronic infectious disease caused by the human immunodeficiency virus (HIV), and the current treatment for the disease is a combination of highly effective antiretroviral therapy, consisting of a combination of at least three of these drugs to combat HIV infection. However, this therapy only inhibits HIV replication, but does not completely eliminate the virus. Once antiretroviral treatment is interrupted, the patient's disease is prone to relapse. Therefore we need gene editing, which is the process of modifying, adding, deleting and replacing information in the body's DNA in order to modify diseases caused by genetic mutations, which can only be controlled by drugs. It is hoped that in the near future, gene editing technology will revolutionise the treatment of cancer and genetic diseases, thanks to the work of scientists.

Ethical Research on Human Gene Editing

The development of biotechnology will inevitably increase its application in medical and health care. Since the experiment of Huang Jun and his team in 2015, there have been a number of biomedical laboratories trying to apply gene editing technology to human therapy. This has aroused great controversy in international academic circles. Many people believe that the misuse of gene editing will not only destroy the existing ethical construction of society, but also affect the ability of human beings to adapt to the future environment. However, this paper believes that gene editing technology can be gradually applied to human body after the government first makes relevant policy preparations and technological breakthroughs are achieved in epigenetic editing. It is hoped that this paper can dispel the excessive concerns of those who oppose human gene editing and provide constructive suggestions on the development of this technologys humanbody application.

Gene-Editing Technology in Lung Cancer: Models and Therapies

Cancer is a disease that develops when the body's cells expand unchecked. Lung cancer is a general term for cancer that first appears in the lungs. The most common cancer that is lethal is lung cancer. Lung cancer is the commonly diagnosed cancer diagnosed between both female and male in the US, only behind certain forms of skin cancer. Lung cancer is most often caused by smoking. Additionally, smoking other tobacco products (including pipes or cigars), breathing secondhand smoke, being linked to radon or radon at work or home, and lung cancer risk may be increased by having a history of the disease in your family. Furthermore, new developments in genome editing technology have dramatically increased the possibility of curing cancer at its origin. This article discusses potential gene editing methods for treating lung cancer and systematically identifies the locations of common gene changes in the condition.

Rapid detection and genotyping of SARS-CoV-2 Omicron BA.4/5 variants using a RT-PCR and CRISPR-Cas12a-based assay

To establish a rapid detection and genotyping method for SARS-CoV-2 Omicron BA.4/5 variants using CRISPPR-Cas12a gene editing technology. We combined reverse transcription-polymerase chain reaction (RT-PCR) and CRISPR gene editing technology and designed a specific CRISPPR RNA (crRNA) with suboptimal protospacer adjacent motifs (PAM) for rapid detection and genotyping of SARS- CoV-2 Omicron BA.4/5 variants. The performance of this RT- PCR/ CRISPPR-Cas12a assay was evaluated using 43 clinical samples of patients infected by wild-type SARS-CoV-2 and the Alpha, Beta, Delta, Omicron BA. 1 and BA. 4/5 variants and 20 SARS- CoV- 2-negative clinical samples infected with 11 respiratory pathogens. With Sanger sequencing method as the gold standard, the specificity, sensitivity, concordance (Kappa) and area under the ROC curve (AUC) of RT-PCR/CRISPPR-Cas12a assay were calculated. This assay was capable of rapid and specific detection of SARS- CoV-2 Omicron BA.4/5 variant within 30 min with the lowest detection limit of 10 copies/μL, and no cross-reaction was observed in SARS-CoV-2-negative clinical samples infected with 11 common respiratory pathogens. The two Omicron BA.4/5 specific crRNAs (crRNA-1 and crRNA-2) allowed the assay to accurately distinguish Omicron BA.4/5 from BA.1 sublineage and other major SARS-CoV-2 variants of concern. For detection of SARS-CoV-2 Omicron BA.4/5 variants, the sensitivity of the established assay using crRNA-1 and crRNA-2 was 97.83% and 100% with specificity of 100% and AUC of 0.998 and 1.000, respectively, and their concordance rate with Sanger sequencing method was 92.83% and 96.41%, respectively. By combining RT-PCR and CRISPPR-Cas12a gene editing technology, we successfully developed a new method for rapid detection and identification of SARS-CoV-2 Omicron BA.4/5 variants with a high sensitivity, specificity and reproducibility, which allows rapid detection and genotyping of SARS- CoV-2 variants and monitoring of the emerging variants and their dissemination.

Development of Medicinal Products Based on Gene-Editing Technology: Regulatory Practices

Somatic cell genome-editing systems are the most recent gene therapy technology to treat patients with monogenic hereditary cancer or HIV. Gene editing allows for changing or completely removing a defective gene with regularly interspaced short palindromic repeat (CRISPR), zinc-finger nuclease (ZFN), and transcription activator-like effector nuclease (TALEN) systems.The aim of the study was to analyse the existing international experience and regulatory requirements relating to the development of medicinal products based on genome editing of postnatal somatic cells.This article describes the mechanism of action of CRISPR, ZFN, and TALEN systems and compares their advantages and disadvantages. Regulatory and legislative authorities should take a special approach to the development, manufacture, and assessment of medicinal products based on genome editing, as well as to the ethical aspects of their use. Current requirements and recommendations for the development of medicinal products based on genome editing are mostly limited to the need to evaluate the risks of off-target effects and late-onset adverse events and the possibility to adapt clinical trial design to surrogate endpoints, exclude healthy volunteers and comparison groups, and select initial doses for clinical trials based on scientific data. Thus, a regulatory approach should also be developed for the marketing authorisation of medicinal products based on genome-editing systems.

Natural and Engineered Resistance Mechanisms in Plants against Phytoviruses.

Plant viruses, as obligate intracellular parasites, rely exclusively on host machinery to complete their life cycle. Whether a virus is pathogenic or not depends on the balance between the mechanisms used by both plants and viruses during the intense encounter. Antiviral defence mechanisms in plants can be of two types, i.e., natural resistance and engineered resistance. Innate immunity, RNA silencing, translational repression, autophagy-mediated degradation, and resistance to virus movement are the possible natural defence mechanisms against viruses in plants, whereas engineered resistance includes pathogen-derived resistance along with gene editing technologies. The incorporation of various resistance genes through breeding programmes, along with gene editing tools such as CRISPR/Cas technologies, holds great promise in developing virus-resistant plants. In this review, different resistance mechanisms against viruses in plants along with reported resistance genes in major vegetable crops are discussed.

Advances In the Application of CRISPR/Cas9 Gene Editing Technology in Pig Breeding and Production

China is the richest pig breeding resource in the world. According to different conditions in different regions, pigs with different excellent traits were bred. Most pig breeds have the advantages of high fat content and strong environmental adaptability, but have the disadvantages of low lean meat rate and poor meat quality. Some pig breeds have poor resistance to disease and are prone to widespread infection. Only crossing with western pig breeds will introduce unfavorable genes, resulting in a sharp decline in the number of local pig breeds, but there are also shortcomings such as long crossing time. Gene editing (GE) technology has become a new way to regulate pig breeding. In recent years, with the exploration of the regular clustering structure and mechanism of the short palindrome repeat/CRISPR associated protein 9 (CRISPR/Cas9) system, GE technology based on CRISPR/Cas9 can improve lean meat rate and obtain disease resistance by knocking out specific genes and enhancing specific gene expression. This technology can also change the reproductive organs of domestic pigs to control their sex and improve the quality of meat. This article presents the application of CRISPR/Cas9 GE technology in pig breeding and production.