Prime editing

一行要約

Prime editing は Anzalone et al. Nature 2019 で開発された search-and-replace 型 genome editing で、Cas9 nickase + reverse transcriptase fusion + prime editing guide RNA (pegRNA) を用いて DSB を作らずに任意の small substitution / insertion / deletion (約50-100 bp) を programmable に導入。Base editing が C→T / A→G に限定されるのに対し、prime editing は all 12 SNV + indel を扱え、CRISPR/Cas9 の DSB 由来 toxicity / off-target / chromosomal rearrangement リスクを回避。oncology では driver mutation precise modeling, oncogenic mutation correction, CAR-T 同種化 (HLA / TCR knockout + transgene knock-in 同時) 研究が活発。

原理

(1) prime editor protein = Cas9 H840A nickase + engineered MMLV reverse transcriptase。(2) pegRNA = sgRNA spacer + scaffold + reverse transcription template (RT template) + primer binding site (PBS)。(3) prime editor が標的 DNA を nick → flap が PBS と hybridize → RT template を逆転写 → 改変配列を含む 3’ flap 生成 → resolution / DNA repair で改変配列が integrate。(4) PE3 / PE3b: 反対鎖 nick で edit efficiency 向上。(5) PE5max = epegRNA (3’ end 安定化) + engineered editor で efficiency / specificity 改善。(6) twinPE / GRAND: 2 pegRNA で large insertion (kbp scale) 可能。

主要エビデンス / 適用領域

• Driver mutation modeling: KRAS G12C / G12D, EGFR L858R / T790M, p53 R175H 等を isogenic cell line で precise install
• Oncogenic mutation correction: T790M / C797S を correct する therapeutic potential 研究
• CAR-T 同種化: HLA class I knockout + CD52 deletion + CAR transgene knock-in を multi-pegRNA で同時実装
• In vivo prime editing: AAV / LNP / VLP 送達で liver / muscle / retina の preclinical 効果 (oncology in vivo はまだ rare)
• Disease modeling: 1 塩基変異の点滅型 disease model (Alagille, Tay-Sachs 等)、cancer susceptibility allele 機能解析

適用分野と限界

• 強み: DSB なし → off-target / chromosomal rearrangement / p53 activation 低減、12 SNV + small indel programmable、precise (homology arm 設計より直接的)
• 限界: efficiency が CRISPR/Cas9 より低い (5-30% typical, PE5max で改善)、large insertion (>kbp) は twinPE 等の特殊設計、in vivo delivery は LNP / AAV / VLP 検討中、pegRNA 設計が complicated (PBS / RT template length 最適化要)、long-term off-target / RT template-derived random insertion の monitoring 必要

Open Questions

• Efficiency 向上: PE6 / PE7 等 next-gen prime editor 開発
• In vivo delivery: LNP / VLP / AAV の比較と clinical pathway
• Therapeutic application: cancer mutation correction の clinical translation
• CAR-T 同種化: prime editing による universal CAR-T 製品開発
• Large rearrangement (chromosomal recombination, gene insertion >10 kbp) の安定 protocol

関連エンティティ

• 補完手法: CRISPR-screen / Base-editing / scRNA-seq / Multiome / Lineage-tracing
• 関連薬剤: CAR-T-therapy / TCR-T-therapy / KRAS-G12C-inhibitor / EGFR-TKI
• 関連遺伝子: KRAS / EGFR / TP53 / ALK
• 関連概念: Oncogene-addiction / EGFR-T790M-resistance / EGFR-C797S-resistance

関連論文

直近 30 件のみ表示中 (関連 177 件)。 全件は MOC ページ または Obsidian (source Markdown) から辿れます。

Basic-Brain metastasis

• Liu et al. CancerDiscov 2026 — Targeting the lipid metabolism proteins FASN and GPAM in alveolar type II cells decreases lung metastasis
• Liu et al. CancerPathogTher 2024 — Experimental models for cancer brain metastasis

Basic-Extracellular vesicles

• Kudo et al. CellMetab 2022 — Secreted phospholipase A2 modifies extracellular vesicles and accelerates B cell lymphoma
• Mugoni et al. JExtracellBiol 2023 — Integrating extracellular vesicle and circulating cell-free DNA analysis using a single plasma aliquot improves the
• De et al. JExtracellVesicles 2026 — Characterizing the fate of anti-CS1 nanobody displaying extracellular vesicles in multiple myeloma
• Son et al. NatCommun 2025 — Fusogenic lipid nanoparticles for rapid delivery of large therapeutic molecules to exosomes
• Ortmann et al. SciRep 2024 — Large extracellular vesicle (EV) and neutrophil extracellular trap (NET) interaction captured in vivo during systemic

Basic-Immunotherapy

• Zhang et al. Cell 2026 — A convergent uPAR-positive tumor ecosystem creates broad vulnerability to CAR t cell therapy
• Pires et al. NatMater 2026 — IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer
• Puig-Saus et al. Nature 2023 — Neoantigen-targeted CD8+ T cell responses with PD-1 blockade therapy

Basic-Liquid-biopsy

• Wang et al. ClinTranslMed 2025 — Cell-free epigenomes enhanced fragmentomics-based model for early detection of lung cancer

Basic-Neutrophil and myeloid cell

• Ponessa et al. CellRep 2026 — Alveolar macrophages inhibit emphysematous pathology via expression of carbonic anhydrase 4
• Gutierrez et al. Cells 2022 — Human neutrophils generate extracellular vesicles that modulate their functional responses
• Wishart et al. CurrProtoc 2023 — Isolation of mouse neutrophils
• Wang et al. NatCancer 2026 — CD300ld on pathologically activated neutrophils promotes tumor immune suppression by binding phosphatidylserine on CD8
• Chen et al. SSRN 2024 — Circulating cell-in-cell tumor cells with neutrophils signature are hematogenous metastasis-initiating cells
• Kuang et al. STARProtoc 2023 — Protocol for density gradient neutrophil isolation and flow cytometry-based characterization from human peripheral blood
• Poli et al. STARProtoc 2023 — Quantitative cytofluorimetric analysis of mouse neutrophil extracellular traps

Basic-Oncogene and genetics

• Stein et al. CancerDiscov 2023 — LKB1-dependent regulation of TPI1 creates a divergent metabolic liability between human and mouse lung adenocarcinoma

Basic-Others

• Zhou et al. NatBiotechnol 2026 — Digital twins of ex vivo human lungs enable accurate and personalized evaluation of therapeutic efficacy
• Jiang et al. NatNanotechnol 2026 — Efficient prime editing in vivo and in vitro using lipid nanoparticles
• Kang et al. NatRevClinOncol 2026 — Translating ferroptosis into oncology challenges, opportunities and future directions
• Turner et al. NatRevImmunol 2026 — RNA-binding proteins and ribonucleoproteins as determinants of immunity

Basic-Tumor microenvironment

• Nigri et al. CancerDiscov 2026 — Myofibroblasts induce neuroplasticity to promote pancreatic inflammation and cancer progression
• Reyes et al. Cell 2026 — Oncogenic and tumor-suppressive forces converge on a progenitor niche at the benign-to-malignant transition

Clinical-Novel modalities

• Kubo et al. BioProtocol 2026 — Cite as kubo, N. et al. (2026). A feeder cell-free system for chimeric antigen receptor gene transduction into
• Moreno et al. LancetRegHealthEur 2026 — Advances in cell therapy for solid tumours european perspective and future directions

Clinical-Others

• Rohrberg et al. JImmunotherCancer 2026 — First-in-human study of lomvastomig, a PD-1-TIM-3 bispecific antibody, in patients with advanced andor metastatic solid

Clinical-Respiratory diseases

• Rosenfield et al. NEnglJMed 2026 — Ultrasound-facilitated, catheter-directed fibrinolysis for acute pulmonary embolism

Clinical-SCLC

• Khurshid et al. JClinOncol 2023 — Systemic therapy for small-cell lung cancer ASCO-ontario health (cancer care ontario) guideline

合成 (Outputs)

• kras-g12c-crispr-modality-synthesis