---
title: "CRISPR-Cas9 advances: engineered Cas variants, delivery strategies, and therapeutic applications f"
id: "pubmed-42758354"
canonical_url: "https://medichelpline.com/clinical-feed/pubmed-42758354"
content_type: "clinical_feed_article"
specialty: "Pharmacology"
source_name: "PubMed / NCBI"
source_url: "https://pubmed.ncbi.nlm.nih.gov/42758354/"
doi: "10.1007/s11033-026-12771-1"
published_at: "2026-09-18T00:00:00.000Z"
evidence_level: "Journal Article"
license: "CC-BY-NC-4.0 / Informational Use"
---
# CRISPR-Cas9 advances: engineered Cas variants, delivery strategies, and therapeutic applications f
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/pubmed-42758354
- **Specialty:** [Pharmacology](https://medichelpline.com/clinical-feed/pharmacology.md)
- **Primary Source:** PubMed / NCBI
- **Source URL:** [Original Journal Publication](https://pubmed.ncbi.nlm.nih.gov/42758354/)
- **DOI:** [10.1007/s11033-026-12771-1](https://doi.org/10.1007%2Fs11033-026-12771-1)
- **Published At:** 2026-09-18T00:00:00.000Z
- **Evidence Rating:** Journal Article
## Executive GIST (TL;DR)
- The review summarizes recent progress in **CRISPR-Cas9** genome editing, highlighting its precision, efficiency, and simplicity derived from a bacterial adaptive immune system. - Novel **Cas** orthologs and engineered **Cas9** variants have been developed to broaden target range, increase fidelity, and reduce off-target effects. - Optimization of single-guide RNAs (**sgRNAs**) has improved target binding, stability, and complex formation with Cas9, boosting overall editing performance. - Next-generation editors such as **base editors** and **prime editors** permit precise single-nucleotide changes and small insertions/deletions without making double-stranded DNA breaks. - Advanced **delivery systems** — including viral vectors, lipid nanoparticles, and ribonucleoprotein electroporation — support efficient editing in in vitro, ex vivo, and in vivo models, and enable primary cell engineering. - Clinical and translational milestones include therapeutic progress in monogenic conditions such as **sickle cell disease**, **β-thalassemia**, and **cystic fibrosis**, with reports of long-term clinical benefits. - In diabetes research, CRISPR-Cas9 has improved disease modeling, clarified molecular pathways in glucose homeostasis, and opened avenues for cellular and gene-based therapies. - The review emphasizes breakthroughs in editing of primary cells and development of translational models for genetic disorders and diabetes mellitus. - Details such as specific experimental protocols, quantitative outcomes, and comprehensive clinical trial data were not fully reported in the abstract and would be found in the full text and cited references.
## Clinical Analysis & Structured Key Points
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Affiliations Expand ### Affiliations * 1 Department of Pharmacy, Gopalganj Science and Technology University, Gopalganj, Dhaka, 8105, Bangladesh. * 2 National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China. * 3 Department of Genetic Engineering and Biotechnology, Faculty of Health and Life Sciences, Daffodil International University, Dhaka, 1216, Bangladesh. * 4 Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. * 5 Department of Pharmacy, Pabna University of Science and Technology, Pabna, 6600, Bangladesh. * 6 Department of Haematology, Rajshahi Medical College, Rajshahi, Bangladesh. * 7 Department of Pharmacy, School of Pharmacy and Public Health, Independent University, Bangladesh, Dhaka, 1229, Bangladesh. shafayat@iub.edu.bd. * 8 School of Biosciences, University of Birmingham, England, Birmingham, United Kingdom. shafayat@iub.edu.bd. * PMID: **42758354** * DOI: [ 10.1007/s11033-026-12771-1 ](https://doi.org/10.1007/s11033-026-12771-1) Item in Clipboard Review # CRISPR-Cas9 genome editing: technological advances, delivery strategies and precision engineering of primary cells for therapeutic application in genetic disorders and diabetes mellitus Md Nur Islam et al. Mol Biol Rep. 2026. Show details Display options Display options Format Abstract PubMed PMID Mol Biol Rep Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Mol+Biol+Rep%22%5Bjour%5D&sort=date&sort_order=desc) * [ Search in NLM Catalog ](https://www.ncbi.nlm.nih.gov/nlmcatalog?term=%22Mol+Biol+Rep%22%5BTitle+Abbreviation%5D) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42758354/) . 2026 Sep 18;53(1):1589. doi: 10.1007/s11033-026-12771-1. ### Authors [Md Nur Islam](https://pubmed.ncbi.nlm.nih.gov/?term=Islam+MN&cauthor_id=42758354)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42758354/#short-view-affiliation-1 "Department of Pharmacy, Gopalganj Science and Technology University, Gopalganj, Dhaka, 8105, Bangladesh.")[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42758354/#short-view-affiliation-2 "National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China."), [Md Mahmudul Islam](https://pubmed.ncbi.nlm.nih.gov/?term=Islam+MM&cauthor_id=42758354)[ 3 ](https://pubmed.ncbi.nlm.nih.gov/42758354/#short-view-affiliation-3 "Department of Genetic Engineering and Biotechnology, Faculty of Health and Life Sciences, Daffodil International University, Dhaka, 1216, Bangladesh."), [Han Feng](https://pubmed.ncbi.nlm.nih.gov/?term=Feng+H&cauthor_id=42758354)[ 4 ](https://pubmed.ncbi.nlm.nih.gov/42758354/#short-view-affiliation-4 "Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China."), [Sm Faysal Bellah](https://pubmed.ncbi.nlm.nih.gov/?term=Bellah+SF&cauthor_id=42758354)[ 5 ](https://pubmed.ncbi.nlm.nih.gov/42758354/#short-view-affiliation-5 "Department of Pharmacy, Pabna University of Science and Technology, Pabna, 6600, Bangladesh."), [M Morsed Zaman Miah](https://pubmed.ncbi.nlm.nih.gov/?term=Zaman+Miah+MM&cauthor_id=42758354)[ 6 ](https://pubmed.ncbi.nlm.nih.gov/42758354/#short-view-affiliation-6 "Department of Haematology, Rajshahi Medical College, Rajshahi, Bangladesh."), [Md Shafayat Hossain](https://pubmed.ncbi.nlm.nih.gov/?term=Hossain+MS&cauthor_id=42758354)[ 7 ](https://pubmed.ncbi.nlm.nih.gov/42758354/#short-view-affiliation-7 "Department of Pharmacy, School of Pharmacy and Public Health, Independent University, Bangladesh, Dhaka, 1229, Bangladesh. shafayat@iub.edu.bd.")[ 8 ](https://pubmed.ncbi.nlm.nih.gov/42758354/#short-view-affiliation-8 "School of Biosciences, University of Birmingham, England, Birmingham, United Kingdom. shafayat@iub.edu.bd.") ### Affiliations * 1 Department of Pharmacy, Gopalganj Science and Technology University, Gopalganj, Dhaka, 8105, Bangladesh. * 2 National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China. * 3 Department of Genetic Engineering and Biotechnology, Faculty of Health and Life Sciences, Daffodil International University, Dhaka, 1216, Bangladesh. * 4 Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. * 5 Department of Pharmacy, Pabna University of Science and Technology, Pabna, 6600, Bangladesh. * 6 Department of Haematology, Rajshahi Medical College, Rajshahi, Bangladesh. * 7 Department of Pharmacy, School of Pharmacy and Public Health, Independent University, Bangladesh, Dhaka, 1229, Bangladesh. shafayat@iub.edu.bd. * 8 School of Biosciences, University of Birmingham, England, Birmingham, United Kingdom. shafayat@iub.edu.bd. * PMID: **42758354** * DOI: [ 10.1007/s11033-026-12771-1 ](https://doi.org/10.1007/s11033-026-12771-1) Item in Clipboard Cite Display options Display options Format Abstract PubMed PMID ## Abstract The CRISPR-Cas9 system has revolutionized modern life sciences, driving a paradigm shift in biomedical research and becoming an indispensable tool in molecular biology due to its remarkable precision, efficiency, and simplicity. Originating from a bacterial adaptive immune mechanism, CRISPR-Cas9 has evolved rapidly, providing a versatile framework for manipulating genetic material and addressing a wide spectrum of human diseases. Recent progress includes the discovery of novel Cas orthologs and the rational engineering of Cas9 variants to enhance editing fidelity, broaden target range, and minimize off-target effects. Structural and functional optimization of single-guide RNAs (sgRNAs) has further improved target binding affinity, stability, and Cas9-sgRNA complex formation, thereby increasing overall editing performance. The development of high-fidelity Cas9 derivatives and next-generation platforms such as base editors and prime editors has enabled precise single-nucleotide substitutions and small insertions or deletions without generating double-stranded DNA breaks. Advanced delivery systems-including viral vectors, lipid nanoparticles, and ribonucleoprotein electroporation-have facilitated efficient CRISPR-mediated editing across in vitro, ex vivo, and in vivo models. Remarkable therapeutic milestones have been achieved in treating monogenic disorders such as sickle cell disease, β-thalassemia, and cystic fibrosis, where long-term clinical benefits have been documented. Furthermore, CRISPR-Cas9 technology is redefining diabetes research by enabling precise modeling of disease mechanisms, uncovering molecular pathways involved in glucose homeostasis, and opening new avenues for cellular and gene-based therapies. This review highlights recent advances in CRISPR-Cas9-mediated genome editing, emphasizing breakthroughs in primary cell editing and the development of translational models for genetic diseases and diabetes mellitus. **Keywords:** CRISPR-Cas9; Cas9 variants; Diabetes mellitus; Genetic diseases; Genome editing. © 2026. The Author(s), under exclusive licence to Springer Nature B.V. [PubMed Disclaimer](https://pubmed.ncbi.nlm.nih.gov/disclaimer/) ## Conflict of interest statement Declarations. Ethics approval and consent to participate: This is a review research study and is exempt from institutional review board approval. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests. ## References 1. 1. Chaudhary A, Kumar V (2025) Rare diseases: a comprehensive literature review and future directions. J Rare Dis 4(1):33 - [DOI](https://doi.org/10.1007/s44162-025-00099-6) 2. 1. Knott GJ, Doudna JA (2018) CRISPR-Cas guides the future of genetic engineering. Science 361(6405):866–869 - [PubMed](https://pubmed.ncbi.nlm.nih.gov/30166482/) - [PMC](https://pmc.ncbi.nlm.nih.gov/articles/6455913/) - [DOI](https://doi.org/10.1126/science.aat5011) 3. 1. Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (2012) A programmable dual-RNA–guided DNA endonuclease in adaptive bacterial immunity. Science 337(6096):816–821 - [PubMed](https://pubmed.ncbi.nlm.nih.gov/22745249/) - [PMC](https://pmc.ncbi.nlm.nih.gov/articles/6286148/) - [DOI](https://doi.org/10.1126/science.1225829) 4. 1. Wu Y, Liang D, Wang Y, Bai M, Tang W, Bao S, Yan Z, Li D, Li J (2013) Correction of a genetic disease in mouse via use of CRISPR-Cas9. Cell Stem Cell 13(6):659–662 - [PubMed](https://pubmed.ncbi.nlm.nih.gov/24315440/) - [DOI](https://doi.org/10.1016/j.stem.2013.10.016) 5. 1. Schwank G, Koo B-K, Sasselli V, Dekkers JF, Heo I, Demircan T, Sasaki N, Boymans S, Cuppen E, Van Der Ent CK (2013) Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients. Cell Stem Cell 13(6):653–658 - [PubMed](https://pubmed.ncbi.nlm.nih.gov/24315439/) - [DOI](https://doi.org/10.1016/j.stem.2013.11.002) Show all 100 references ## Publication types * Review Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Review%22%5Bpt%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Review) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42758354/) ## MeSH terms * Animals Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Animals%22%5BMeSH%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Animals) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42758354/) * CRISPR-Cas Systems* / genetics Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22CRISPR-Cas+Systems%2Fgenetics%22%5BMAJR%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=CRISPR-Cas+Systems) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42758354/) * Diabetes Mellitus* / genetics Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Diabetes+Mellitus%2Fgenetics%22%5BMAJR%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Diabetes+Mellitus) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42758354/) * Diabetes Mellitus* / therapy Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Diabetes+Mellitus%2Ftherapy%22%5BMAJR%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Diabetes+Mellitus) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42758354/) * Gene Editing* / methods Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Gene+Editing%2Fmethods%22%5BMAJR%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Gene+Editing) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42758354/) * Genetic Diseases, Inborn* / genetics Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Genetic+Diseases%2C+Inborn%2Fgenetics%22%5BMAJR%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Genetic+Diseases%2C+Inborn) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42758354/) * Genetic Diseases, Inborn* / therapy Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Genetic+Disease
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