MutSα and MutSβ as size-dependent cellular determinants for prime editing in human embryonic stem cells

Park, Ju-Chan; Kim, Yun-Jeong; Han, Jun; Kim, Da-Yeon; Park, Mihn Jeong; Kim, Jumee; Jang, Hyeon-Ki; Bae, Sangsu; Cha, Hyuk‐Jin

doi:10.1016/j.omtn.2023.05.015

Cited by 10 publications

(3 citation statements)

References 51 publications

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“…Recent research has drawn attention to the impact of the DNA damage response and repair pathways on the e cacy of precise genome editing tools like CBE and PE in hPSCs 36,43 . Informed by these ndings, we've demonstrated in the present study that concurrent inhibition of both the DNA damage response and DNA repair pathways represents a straightforward strategy to enhance editing outcomes.…”

Section: Discussionmentioning

confidence: 99%

Enhancing Precise Genome Editing in Human Pluripotent Stem Cells through Dual Inhibition of DNA Damage Response and Repair Pathways

Park

Kim

Kang

et al. 2023

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Precise genome editing is crucial for establishing isogenic human disease models and ex vivo stem cell therapy from the patient-derived human pluripotent stem cells (hPSCs). Unlike Cas9-mediated knock-in, cytosine base editor (CBE) and prime editor (PE) achieve the desirable gene correction without inducing DNA double strand breaks. However, hPSCs possess highly active DNA repair systems and are particularly susceptible to p53-dependent cell death. These unique characteristics impede the efficiency of gene editing in hPSCs. Here, we demonstrate that dual inhibition of p53-mediated cell death and distinct activation of the DNA damage repair system upon DNA damage by CBE or PE additively enhanced editing efficiency in hPSCs. The BE4stem system comprised of dominant negative p53 (p53DD) and three UNG inhibitor (UGI), engineered to specifically diminish base excision repair (BER), improved CBE efficiency in hPSCs. Addition of dominant negative MLH1 to inhibit mismatch repair activity and p53DD in the conventional PE system also significantly enhanced PE efficiency in hPSCs. Thus, combined inhibition of the unique cellular cascades engaged in hPSCs upon gene editing could significantly enhance precise genome editing in these cells.

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Section: Discussionmentioning

confidence: 99%

Enhancing Precise Genome Editing in Human Pluripotent Stem Cells through Dual Inhibition of DNA Damage Response and Repair Pathways

Park

Kim

Kang

et al. 2023

Preprint

Self Cite

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show abstract

“…In summary, the findings of Park et al. 4 provide valuable insights into the inhibitory role of MutSα and MutSβ complexes in PE outcomes. A better understanding of the inhibitory role of MMR can lead to strategies for overcoming the limitations in PE, opening new possibilities for precise and reliable genetic modifications.…”

mentioning

confidence: 89%

“…To address this critical roadblock, in a recent study, Park et al. 4 investigated the role of the MMR complexes MutSα (MSH2-MSH6 subunits) and MutSβ (MSH2-MSH3 subunits) in human embryonic stem cells (hESCs). Because MSH2 , MSH3 , and MSH6 gene expression is higher in hESCs compared to differentiated cell derivatives, the authors hypothesized that MutSα and MutSβ expression hinders the efficiency of PE in hESCs.…”

mentioning

confidence: 99%