Enhanced Calvarial Bone Repair Using ASCs Engineered with RNA‐Guided Split dCas12a System that Co‐Activates Sox 5, Sox6, and Long Non‐Coding RNA H19
Healing of large calvarial bone defects remains challenging. An RNA‐guided Split dCas12a system is previously harnessed to activate long non‐coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow‐derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic...
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| Published in | Small (Weinheim an der Bergstrasse, Germany) Vol. 20; no. 21; pp. e2306612 - n/a |
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| Main Authors | , , , , , , , , , , , , |
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| Language | English |
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01.05.2024
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| Abstract | Healing of large calvarial bone defects remains challenging. An RNA‐guided Split dCas12a system is previously harnessed to activate long non‐coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow‐derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose‐derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However, H19 activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co‐activate other chondroinductive transcription factors (Sox5, Sox6, and Sox9) to synergistically potentiate differentiation. It is found that co‐activation of H19/Sox5/Sox6 in ASCs elicited more potent chondrogenic differentiation than activation of Sox5/Sox6/Sox9 or H19 alone. Co‐activating H19/Sox5/Sox6 in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration.
A new Split dCas12a activator is designed for delivery into ASCs to co‐activate long non‐coding RNA H19, Sox5, and Sox6. The split dCas12a spontaneously dimerizes in ASCs and activates H19, Sox5, and Sox6, hence enhancing chondrogenesis, suppressing adipogenesis, and augmenting calvarial bone healing in rats. |
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| AbstractList | Healing of large calvarial bone defects remains challenging. An RNA‐guided Split dCas12a system is previously harnessed to activate long non‐coding RNA
H19
(lncRNA
H19
, referred to as
H19
thereafter) in bone marrow‐derived mesenchymal stem cells (BMSCs).
H19
activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose‐derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However,
H19
activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co‐activate other chondroinductive transcription factors (
Sox5
,
Sox6
, and
Sox9
) to synergistically potentiate differentiation. It is found that co‐activation of
H19
/
Sox5
/
Sox6
in ASCs elicited more potent chondrogenic differentiation than activation of
Sox5
/
Sox6/Sox9
or
H19
alone. Co‐activating
H19
/
Sox5/Sox6
in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration. Healing of large calvarial bone defects remains challenging. An RNA-guided Split dCas12a system is previously harnessed to activate long non-coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow-derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose-derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However, H19 activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co-activate other chondroinductive transcription factors (Sox5, Sox6, and Sox9) to synergistically potentiate differentiation. It is found that co-activation of H19/Sox5/Sox6 in ASCs elicited more potent chondrogenic differentiation than activation of Sox5/Sox6/Sox9 or H19 alone. Co-activating H19/Sox5/Sox6 in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration.Healing of large calvarial bone defects remains challenging. An RNA-guided Split dCas12a system is previously harnessed to activate long non-coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow-derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose-derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However, H19 activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co-activate other chondroinductive transcription factors (Sox5, Sox6, and Sox9) to synergistically potentiate differentiation. It is found that co-activation of H19/Sox5/Sox6 in ASCs elicited more potent chondrogenic differentiation than activation of Sox5/Sox6/Sox9 or H19 alone. Co-activating H19/Sox5/Sox6 in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration. Healing of large calvarial bone defects remains challenging. An RNA‐guided Split dCas12a system is previously harnessed to activate long non‐coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow‐derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose‐derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However, H19 activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co‐activate other chondroinductive transcription factors (Sox5, Sox6, and Sox9) to synergistically potentiate differentiation. It is found that co‐activation of H19/Sox5/Sox6 in ASCs elicited more potent chondrogenic differentiation than activation of Sox5/Sox6/Sox9 or H19 alone. Co‐activating H19/Sox5/Sox6 in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration. Healing of large calvarial bone defects remains challenging. An RNA-guided Split dCas12a system is previously harnessed to activate long non-coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow-derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose-derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However, H19 activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co-activate other chondroinductive transcription factors (Sox5, Sox6, and Sox9) to synergistically potentiate differentiation. It is found that co-activation of H19/Sox5/Sox6 in ASCs elicited more potent chondrogenic differentiation than activation of Sox5/Sox6/Sox9 or H19 alone. Co-activating H19/Sox5/Sox6 in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration. Healing of large calvarial bone defects remains challenging. An RNA‐guided Split dCas12a system is previously harnessed to activate long non‐coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow‐derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose‐derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However, H19 activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co‐activate other chondroinductive transcription factors (Sox5, Sox6, and Sox9) to synergistically potentiate differentiation. It is found that co‐activation of H19/Sox5/Sox6 in ASCs elicited more potent chondrogenic differentiation than activation of Sox5/Sox6/Sox9 or H19 alone. Co‐activating H19/Sox5/Sox6 in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration. A new Split dCas12a activator is designed for delivery into ASCs to co‐activate long non‐coding RNA H19, Sox5, and Sox6. The split dCas12a spontaneously dimerizes in ASCs and activates H19, Sox5, and Sox6, hence enhancing chondrogenesis, suppressing adipogenesis, and augmenting calvarial bone healing in rats. |
| Author | Dang, Quyen Thuc Lee, Shang‐Shung Chang, Chin‐Wei Truong, Vy Anh Chen, Pin‐Hsin Pham, Nam Ngoc Ngo, Dung Kim Thi Hu, Yu‐Chen Truong, Vu Anh Chang, Yi‐Hao Pham, Dang Huu Chang, Yu‐Han Nguyen, Nuong Thi Kieu |
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| Keywords | H19 calvarial bone healing CRISPR activation Split dCas12a Sox6 Sox5 |
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| Snippet | Healing of large calvarial bone defects remains challenging. An RNA‐guided Split dCas12a system is previously harnessed to activate long non‐coding RNA H19... Healing of large calvarial bone defects remains challenging. An RNA‐guided Split dCas12a system is previously harnessed to activate long non‐coding RNA H19... Healing of large calvarial bone defects remains challenging. An RNA-guided Split dCas12a system is previously harnessed to activate long non-coding RNA H19... |
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| SubjectTerms | Adipose Tissue - cytology Animals Bone marrow calvarial bone healing Cell Differentiation Chondrogenesis CRISPR activation Differentiation H19 Healing Humans Mesenchymal Stem Cells - cytology Mesenchymal Stem Cells - metabolism Osteogenesis - genetics Regeneration (physiology) Ribonucleic acid RNA RNA, Long Noncoding - genetics RNA, Long Noncoding - metabolism Skull Sox5 Sox6 SOXD Transcription Factors - genetics SOXD Transcription Factors - metabolism Split dCas12a Stem cells Stem Cells - cytology Stem Cells - metabolism Tissue engineering |
| Title | Enhanced Calvarial Bone Repair Using ASCs Engineered with RNA‐Guided Split dCas12a System that Co‐Activates Sox 5, Sox6, and Long Non‐Coding RNA H19 |
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