Collection Overview
CRISPR-based technologies have transformed biological research by enabling precise, efficient, and versatile manipulation of genetic material across a wide range of organisms, cell types, and experimental systems. This topical collection brings together practical protocols and methodological advances that support the application of CRISPR and related genome engineering tools in fundamental research, disease modelling, therapeutic development, and molecular diagnostics.
The collection covers diverse approaches to genome editing, including CRISPR/Cas9-mediated gene knockout, targeted integration, precise knock-in, base editing, endogenous protein tagging, and epigenome modification. It also encompasses CRISPR interference and activation strategies, multiplex editing, pooled and genome-wide genetic screens, and methods for investigating gene function, regulatory elements, chromatin organisation, and genetic interactions. Particular attention is given to experimental workflows that improve editing efficiency, specificity, validation, and reproducibility, including guide RNA design, vector construction, viral delivery, electroporation, microinjection, and the identification and characterisation of edited cells and organisms.
Contributions span an extensive range of biological models, from mammalian cell lines, human induced pluripotent stem cells, and primary immune and stem cells to mice, zebrafish, Drosophila, Caenorhabditis elegans, insects, plants, fungi, parasites, and other microorganisms. Applications include cancer biology, immunology, infectious disease, gene therapy, developmental biology, metabolism, and functional genomics. The collection also highlights emerging CRISPR-based diagnostic technologies, including nucleic acid detection, portable and point-of-care assays, and high-throughput screening platforms.
By bringing together video-based experimental protocols and methodological innovations, this collection aims to help researchers select, implement, and adapt CRISPR-based techniques to their own experimental questions. It will serve as a practical resource for scientists seeking reliable, reproducible approaches to genome engineering, functional investigation, and the development of next-generation diagnostic and therapeutic technologies.
Articles
Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9
0 Views
•
Cited by 157
•
2015
•
Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System
0 Views
•
Cited by 5
•
2015
•
1Department of Neurosurgery, The University of Texas Health Science Center at Houston, 2Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 3The Senator Lloyd & B. A. Bentsen Center for Stroke Research, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 4Summer Research Program, Office of Educational Programs, The University of Texas Health Science Center at Houston, 5Department of Anesthesiology, Shengjing Hospital, China Medical University, 6Department of Oncology, Renji Hospital, Shanghai Jiaotong University School of Medicine, 7Biology Department, University of West Georgia
QTL Mapping and CRISPR/Cas9 Editing to Identify a Drug Resistance Gene in Toxoplasma gondii
0 Views
•
Cited by 5
•
2017
•
Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex
0 Views
•
Cited by 24
•
2018
•
Rong Li*1, Jinxin Miao*1,2, Zhiqiang Fan1, SeokHwan Song3, Il-keun Kong3,4, Yaohe Wang2,5, Zhongde Wang1
1Department of Animal, Dairy, and Veterinary Sciences, Utah State University, 2National Centre for International Research in Cell and Gene Therapy, Sino-British Research Centre, School of Basic Medical Sciences, Zhengzhou University, 3Department of Animal Science Division of Applied Life Science (BK21 Plus), Gyeongsang National University, 4Institute of Agriculture and Life Science, Gyeongsang National University, 5Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London
Microinjection of CRISPR/Cas9 Protein into Channel Catfish, Ictalurus punctatus, Embryos for Gene Editing
0 Views
•
Cited by 34
•
2018
•
Ahmed Elaswad*1,2, Karim Khalil*1,3, David Cline1, Patrick Page-McCaw4, Wenbiao Chen4, Maximilian Michel5, Roger Cone5, Rex Dunham1
1School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, 2Department of Animal Wealth Development, Faculty of Veterinary Medicine, Suez Canal University, 3Anatomy and Embryology Department, Faculty of Veterinary Medicine, Cairo University, 4Department of Molecular Physiology and Biophysics, Vanderbilt University, 5Life Science Institute, University of Michigan
CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
0 Views
•
Cited by 15
•
2018
•
1Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 2College of Life Sciences, University of Chinese Academy of Sciences, 3School of Life Science and Technology, Shanghai Tech University, 4Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences
Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9
0 Views
•
Cited by 26
•
2018
•
Lorenzo Brunetti*1,2,3, Michael C. Gundry*1,2,4, Ayumi Kitano4, Daisuke Nakada1,2,4, Margaret A. Goodell1,2,4,5
1Stem Cells & Regenerative Medicine Center, Baylor College of Medicine, 2Center for Cell and Gene Therapy, Baylor College of Medicine, 3Centro di Ricerca Emato-Oncologica (CREO), University of Perugia, 4Department of Molecular & Human Genetics, Baylor College of Medicine, 5Texas Children's Hospital & Houston Methodist Hospital
Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration
0 Views
•
Cited by 12
•
2018
•
CRISPR-mediated Loss of Function Analysis in Cerebellar Granule Cells Using In Utero Electroporation-based Gene Transfer
0 Views
•
Cited by 4
•
2018
•
Weijun Feng*1, Lena Herbst*2, Peter Lichter2, Stefan M. Pfister3,4,5, Hai-Kun Liu1, Daisuke Kawauchi3,4
1Division of Molecular Neurogenetics, German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, 2Division of Molecular Genetics, German Cancer Research Center (DKFZ), and German Cancer Consortium (DKTK), 3Hopp-Children's Cancer Center at the NCT Heidelberg (KiTZ), 4Division of Pediatric Neurooncology, German Cancer Research Center (DKFZ), and German Cancer Consortium (DKTK), 5Department of Pediatric Hematology and Oncology, Heidelberg University Hospital
Adeno-Associated Virus-Mediated Delivery of CRISPR for Cardiac Gene Editing in Mice
0 Views
•
Cited by 5
•
2018
•
CRISPR-Mediated Reorganization of Chromatin Loop Structure
0 Views
•
Cited by 7
•
2018
•
Stefanie L. Morgan*1,2, Erin Y. Chang*1, Natasha C. Mariano1, Abel Bermudez3, Nicole L. Arruda4, Fanting Wu5, Yunhai Luo1, Gautam Shankar1, Star K. Huynh1, Chiao-Chain Huang5, Sharon J. Pitteri3, Kevin C. Wang1,2,6
1Department of Dermatology, Program in Epithelial Biology, Stanford University School of Medicine, 2Program in Cancer Biology, Stanford University School of Medicine, 3Canary Center for Cancer Early Detection, Department of Radiology, Stanford University School of Medicine, 4Department of Biology, Bridgewater State University, 5System Biosciences, 6Veterans Affairs Healthcare System
CRISPR Guide RNA Cloning for Mammalian Systems
0 Views
•
Cited by 16
•
2018
•
Sathiji Nageshwaran*1,2, Alejandro Chavez*1,2,3, Nan Cher Yeo1,2, Xiaoge Guo1,2, Alissa Lance-Byrne1, Angela Tung1, James J. Collins1,4,5,6,7, George M. Church1,2
1Wyss Institute for Biologically Inspired Engineering, Harvard University, 2Department of Genetics, Harvard Medical School, 3Department of Pathology, Massachusetts General Hospital, 4Institute for Medical Engineering & Science, Massachusetts Institute of Technology, 5Synthetic Biology Center, Massachusetts Institute of Technology, 6Department of Biological Engineering, Massachusetts Institute of Technology, 7Broad Institute
CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis
0 Views
•
Cited by 7
•
2020
•
Overexpressing Long Noncoding RNAs Using Gene-activating CRISPR
0 Views
•
Cited by 6
•
2019
•
Carl Robert Rankin1, Janet Treger1, Emmanuelle Faure-Kumar2, Jihane Benhammou1, Deborah Anisman-Posner3, Alex Edward Bollinger3, Charalabos Pothoulakis1, David Miguel Padua1
1Vatche and Tamar Manoukian Division of Digestive Diseases, Department of Medicine, University of California, Los Angeles, 2Vatche and Tamar Manoukian Division of Digestive Diseases, Integrated Molecular Technologies (IMT) Core, Department of Medicine, University of California, Los Angeles, 3Division of Hematology and Oncology, Department of Medicine, University of California, Los Angeles
Genome Editing in Mammalian Cell Lines using CRISPR-Cas
0 Views
•
Cited by 5
•
2019
•
Using CRISPR/Cas9 to Knock Out GM-CSF in CAR-T Cells
0 Views
•
Cited by 34
•
2019
•
Pooled CRISPR-Based Genetic Screens in Mammalian Cells
0 Views
•
Cited by 13
•
2019
•
Silencing the Spark: CRISPR/Cas9 Genome Editing in Weakly Electric Fish
0 Views
•
Cited by 7
•
2019
•
Preparing and Injecting Embryos of Culex Mosquitoes to Generate Null Mutations using CRISPR/Cas9
0 Views
•
Cited by 5
•
2020
•
Pupal and Adult Injections for RNAi and CRISPR Gene Editing in Nasonia vitripennis
0 Views
•
Cited by 5
•
2020
•
Embryo Injections for CRISPR-Mediated Mutagenesis in the Ant Harpegnathos saltator
0 Views
•
Cited by 9
•
2021
•
Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors
0 Views
•
Cited by 1
•
2021
•
Production of Human CRISPR-Engineered CAR-T Cells
0 Views
•
Cited by 19
•
2021
•
Embryo Microinjection and Knockout Mutant Identification of CRISPR/Cas9 Genome-Edited Helicoverpa Armigera (Hübner)
0 Views
•
Cited by 4
•
2021
•
1School of Chemical and Environmental Engineering, China University of Mining and Technology, 2State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 3Guangdong Laboratory of Lingnan Modern Agriculture, Shenzhen; Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences
Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis
0 Views
•
Cited by 8
•
2022
•
1Key Laboratory of Sustainable Forest Ecosystem Management - Ministry of Education, Northeast Forestry University, 2Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences
Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a
0 Views
•
Cited by 2
•
2022
•
Min Li*1, Hongkun Qin*1, Yunfei Long1, Meiqin Cheng1, Ling Li1, Aijun Huang2, Nian Wang3, Shuo Duan1
1China-USA Citrus Huanglongbing Joint Laboratory, National Navel Orange Engineering Research Center, Gannan Normal University, 2Department of Biological Sciences, Gannan Normal University, 3Citrus Research and Education Center, Department of Microbiology and Cell Science, University of Florida - Institute of Food and Agricultural Sciences
Mouse In Vivo Placental Targeted CRISPR Manipulation
0 Views
•
Cited by 5
•
2023
•
Generation of Centromere-Associated Protein-E CENP-E-/- Knockout Cell Lines using the CRISPR/Cas9 System
0 Views
•
2023
•
Meng-Fei Xu1,2, Jie Chen1,2, Yue Xu1,2, Jing-Lian Zhang1,2, Yi Zhou1,2, Jie-Jie He1,2, Shan Wu1,2, Ya-Lan Wei3,4, Zhen-Yu She1,2
1Department of Cell Biology and Genetics, The School of Basic Medical Sciences, Fujian Medical University, 2Key Laboratory of Stem Cell Engineering and Regenerative Medicine, Fujian Province University, 3College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, 4Medical Research Center, Fujian Maternity and Child Health Hospital
CRISPR-Cas-mediated Multianalyte Synthetic Urine Biomarker Test for Portable Diagnostics
0 Views
•
Cited by 3
•
2023
•
Audrey E. Van Heest1, Feiyang Deng1, Renee T. Zhao2, Nour Saida Harzallah2,3, Heather E. Fleming3,4, Sangeeta N. Bhatia2,3,4,5,6,7,8, Liangliang Hao1,2,3
1Department of Biomedical Engineering, Boston University, 2Institute for Medical Engineering and Science, Massachusetts Institute of Technology, 3Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, 4Howard Hughes Medical Institute, 5Broad Institute of Massachusetts Institute of Technology and Harvard, 6Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, 7Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, 8Department of Medicine, Brigham and Women’s Hospital and Harvard Medical School
One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster
0 Views
•
Cited by 2
•
2024
•
CRISPR-Based Modular Assembly for High-Throughput Construction of a UAS-cDNA/ORF Plasmid Library
0 Views
•
Cited by 1
•
2024
•
Si Xu*1,2, Mengyu Chen*1,2, Guang Chen1,3, Si Luo4, Wen Xue1,2, Xuelian Liu1,2, Jiwu Wang1,2,3, Ping Wei4
1Clinical Research Institute, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, 2Department of Neurology, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, 3Department of Clinical Laboratory, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, 4Shanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, Shanghai Clinical Center for Diabetes, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine
CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions
0 Views
•
Cited by 3
•
2024
•
Muhammad Abid Sheikh1, Farah Helena Afandi1, Grazia Iannello2, Barbara Corneo2, Bright Starling Emerald3,4,5, Suraiya Anjum Ansari1,4,5
1Department of Biochemistry and Molecular Biology, College of Medicine and Health Sciences, United Arab Emirates University, 2Columbia Stem Cell Initiative, Stem Cell Core, Columbia University Irving Medical Center, 3Department of Anatomy, College of Medicine and Health Sciences, United Arab Emirates University, 4Zayed Center for Health Sciences, United Arab Emirates University, 5ASPIRE Precision Medicine Research Institute Abu Dhabi (PMRI-AD), United Arab Emirates University
Retroviral CRISPR/Cas9-Mediated Gene Targeting for the Study of Th17 Differentiation in Vitro
0 Views
•
Cited by 1
•
2024
•
1Shanghai Immune Therapy Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, 2Zhejiang University-University of Edinburgh Institute (ZJU-UoE Institute), Zhejiang University School of Medicine, Zhejiang University, 3Institute of Immunology and Department of Rheumatology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine
Performing an In Vitro Genome-Wide CRISPR Knockout Screen in Chimeric Antigen Receptor T Cells
0 Views
•
2025
•
1T Cell Engineering, Mayo Clinic, 2Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic, 3Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, 4Center for Regenerative Biotherapeutics, Mayo Clinic, 5Division of Hematology, Mayo Clinic, 6Department of Immunology, Mayo Clinic
Genome Editing in the Yellow Fever Mosquito Aedes aegypti using CRISPR-Cas9
0 Views
•
Cited by 2
•
2025
•
Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes
0 Views
•
2025
•
1Department of Pulmonary Oncology, Zhongnan Hospital of Wuhan University, 2Tumor Precision Diagnosis and Treatment Technology and Translational Medicine, Hubei Engineering Research Center, Zhongnan Hospital of Wuhan University, 3Hubei Key Laboratory of Tumor Biological Behaviors, Zhongnan Hospital of Wuhan University
CRISPR-based Shuttle Cloning: A High-throughput Cloning Method
0 Views
•
2025
•
1Institute of Biochemistry and Molecular Biology, Hengyang Medical School, University of South China, 2Clinical Research Institute, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, 3Department of Clinical Laboratory, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, 4Shanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, Shanghai Clinical Center for Diabetes, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine
On-site DNA Detection of Trypanosomatid Parasites and Nosema ceranae Through Alkaline Lysis Coupled to RPA/CRISPR/Cas12a System
0 Views
•
2025
•
Precise Phage Mutagenesis with NgTET-Assisted CRISPR-Cas Systems
0 Views
•
2025
•
1Max Planck Institute for Terrestrial Microbiology, 2Kavli Institute of Nanoscience, Delft University of Technology, 3Department of Bionanoscience, Delft University of Technology, 4Philipps-Universität Marburg, Department of Chemistry, Center for Synthetic Microbiology (SYNMIKRO), 5Institute of Pharmaceutical Biology and Biotechnology, Department of Pharmacy, Marburg University
Improving CRISPR-Cas9 Screens in CAR T Cells: A Refined Method for Library Preparation
0 Views
•
2026
•
Maider Garnica*1, Patxi San Martin-Uriz*1,2, Paula Rodriguez-Marquez1,2, Maria E. Calleja-Cervantes1,3, Saray Rodriguez-Diaz1, Rebeca Martinez-Turrillas1,2, Mikel Hernaez2,3,4,5, Felipe Prosper1,2,4,6, Juan R. Rodriguez-Madoz1,2,4
1Hemato-Oncology Program, Cima Universidad de Navarra, IdiSNA, 2Centro de Investigacion Biomedica en Red de Cancer (CIBERONC), 3Computational Biology and Translational Genomics Program, Cima Universidad de Navarra, IdiSNA, 4Cancer Center Clinica Universidad de Navarra (CCUN), 5Data Science and Artificial Intelligence Institute (DATAI), Universidad de Navarra, 6Hematology and Cell Therapy Department, Clinica Universidad de Navarra, IdiSNA
One-step Overlapping PCR for Rapid Synthesis of Single-guide RNA DNA Templates for the CRISPR System
0 Views
•
2026
•
1Institute of Cytology and Genetics, School of Basic Medical Sciences, Hengyang Medical School, University of South China, 2Translational Medicine Institute, the First People's Hospital of Chenzhou, Hengyang Medical School, University of South China, 3National Health Commission Key Laboratory of Birth Defect Research and Prevention & Ministry of Education Key Lab of Rare Pediatric Diseases, Institute of Cell Biology and Genetics, School of Basic Medical Sciences, Hengyang Medical School, University of South China
Lentiviral SgRNA Delivery for CRISPR/Cas9 Editing in Cre-Dependent Cas9 Knock-in Primary Mouse Hepatocytes
0 Views
•
2026
•
1Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences, 3Department of Mechanical, Materials, and Manufacturing Engineering, University of Nottingham Ningbo China, 4Advanced Mechanical Engineering, Manufacturing, and Mobility Research Centre, University of Nottingham Ningbo China, 5School of Chinese Materia Medica, Nanjing University of Chinese Medicine
CRISPR/Cas9-Mediated Site-Directed Mutagenesis of Genes in Klebsiella pneumoniae
0 Views
•
2026
•
1Department of Infectious Diseases, the Affiliated Hospital of Guizhou Medical University, 2Guizhou Provincial Key Laboratory for Digestive System Diseases, 3State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, 4Infection and Hepatology Laboratory, the Affiliated Hospital of Guizhou Medical University
Generation and Phenotypic Characterization of a CRISPR/Cas9-Engineered Cracd-Deficient Mouse Model for Post-Myocardial Infarction Remodeling Studies
0 Views
•
2026
•
Shuang Zheng1, Xiang Li1,2, Fei Li1, Xuecong Ren1, Jianxin Chen3, Yunzeng Zou2, Feng Hua Yang1, Yu Xia4, Weijiang Tan1,3
1Guangdong Provincial Biotechnology Research Institute (Guangdong Provincial Laboratory Animals Monitoring Center), 2Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, 3College of Veterinary Medicine, South China Agricultural University, 4Department of Cardiovascular Surgery, Guangdong Provincial Hospital of Chinese Medicine, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine
