Transcriptome-wide detection of RNA cleavage sites revealed tRNA cleavage by target-activated CRISPR-Cas13a effector
Аутори
Kolesnik, MatveiJain, Ishita
Semenova, Ekaterina
Severinov, Konstantin
Остала ауторства
Morić, IvanaĐorđević, Valentina
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
feature of Type VI systems is collateral RNA damage. Specifically, the binding of a target
transcript by Cas13a, charged with cognate CRISPR RNA (crRNA), activates the Cas13a
enzyme, turning it into an active ribonuclease that mediates the cleavage of noncomplementary
RNA molecules. Previously, Cas13a-mediated collateral RNA cleavage
was observed in in vitro experiments and was described to be nonspecific. In Escherichia
coli, targeting of nonessential transcripts by heterologously expressed Leptotrichia shahii
Cas13a enzyme (LshCas13a) leads to cell growth retardation, which was proposed to be a
consequence of collateral degradation of essential cellular transcripts. However, the direct
link between collateral RNA cleavage and cell growth retardation was not established.
Specifically, the products of collateral RNA cleavage mediated by target-activated Cas13a
enzyme were not identified in living cells.
To detect RNA cleavage sites associated with collateral Cas13a activity,... a specific
approach based on high-throughput RNA sequencing was developed. This approach
was successfully applied to detect RNA cleavage sites introduced by target-activated
LshCas13a enzyme in both in vivo and in vitro experiments. In E. coli cells, the targetactivated
LshCas13a enzyme cleaves tRNA molecules within anticodon loops, leading
to protein synthesis inhibition and slowing down cell growth. Additionally, LshCas13amediated
collateral tRNA cleavage indirectly activates cellular ribonucleases encoded by
Type II toxin-antitoxin systems.
Together, the results suggest that the L. shahii Type VI CRISPR-Cas system mediates the
immune response by inhibiting translation through collateral tRNA cleavage.
Кључне речи:
CRISPR-Cas systems / RNA-Seq / Cas13Извор:
5th Belgrade Bioinformatics Conference, 2024, 130-130Издавач:
- Belgrade : Institute of molecular genetics and genetic engineering
Напомена:
- Book of abstracts: 5th Belgrade Bioinformatics Conference, Serbia, Belgrade,17-20 june 2024.
Колекције
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - CONF AU - Kolesnik, Matvei AU - Jain, Ishita AU - Semenova, Ekaterina AU - Severinov, Konstantin PY - 2024 UR - www.belbi.bg.ac.rs UR - https://imagine.imgge.bg.ac.rs/handle/123456789/2470 AB - feature of Type VI systems is collateral RNA damage. Specifically, the binding of a target transcript by Cas13a, charged with cognate CRISPR RNA (crRNA), activates the Cas13a enzyme, turning it into an active ribonuclease that mediates the cleavage of noncomplementary RNA molecules. Previously, Cas13a-mediated collateral RNA cleavage was observed in in vitro experiments and was described to be nonspecific. In Escherichia coli, targeting of nonessential transcripts by heterologously expressed Leptotrichia shahii Cas13a enzyme (LshCas13a) leads to cell growth retardation, which was proposed to be a consequence of collateral degradation of essential cellular transcripts. However, the direct link between collateral RNA cleavage and cell growth retardation was not established. Specifically, the products of collateral RNA cleavage mediated by target-activated Cas13a enzyme were not identified in living cells. To detect RNA cleavage sites associated with collateral Cas13a activity, a specific approach based on high-throughput RNA sequencing was developed. This approach was successfully applied to detect RNA cleavage sites introduced by target-activated LshCas13a enzyme in both in vivo and in vitro experiments. In E. coli cells, the targetactivated LshCas13a enzyme cleaves tRNA molecules within anticodon loops, leading to protein synthesis inhibition and slowing down cell growth. Additionally, LshCas13amediated collateral tRNA cleavage indirectly activates cellular ribonucleases encoded by Type II toxin-antitoxin systems. Together, the results suggest that the L. shahii Type VI CRISPR-Cas system mediates the immune response by inhibiting translation through collateral tRNA cleavage. PB - Belgrade : Institute of molecular genetics and genetic engineering C3 - 5th Belgrade Bioinformatics Conference T1 - Transcriptome-wide detection of RNA cleavage sites revealed tRNA cleavage by target-activated CRISPR-Cas13a effector EP - 130 SP - 130 UR - https://hdl.handle.net/21.15107/rcub_imagine_2470 ER -
@conference{ author = "Kolesnik, Matvei and Jain, Ishita and Semenova, Ekaterina and Severinov, Konstantin", year = "2024", abstract = "feature of Type VI systems is collateral RNA damage. Specifically, the binding of a target transcript by Cas13a, charged with cognate CRISPR RNA (crRNA), activates the Cas13a enzyme, turning it into an active ribonuclease that mediates the cleavage of noncomplementary RNA molecules. Previously, Cas13a-mediated collateral RNA cleavage was observed in in vitro experiments and was described to be nonspecific. In Escherichia coli, targeting of nonessential transcripts by heterologously expressed Leptotrichia shahii Cas13a enzyme (LshCas13a) leads to cell growth retardation, which was proposed to be a consequence of collateral degradation of essential cellular transcripts. However, the direct link between collateral RNA cleavage and cell growth retardation was not established. Specifically, the products of collateral RNA cleavage mediated by target-activated Cas13a enzyme were not identified in living cells. To detect RNA cleavage sites associated with collateral Cas13a activity, a specific approach based on high-throughput RNA sequencing was developed. This approach was successfully applied to detect RNA cleavage sites introduced by target-activated LshCas13a enzyme in both in vivo and in vitro experiments. In E. coli cells, the targetactivated LshCas13a enzyme cleaves tRNA molecules within anticodon loops, leading to protein synthesis inhibition and slowing down cell growth. Additionally, LshCas13amediated collateral tRNA cleavage indirectly activates cellular ribonucleases encoded by Type II toxin-antitoxin systems. Together, the results suggest that the L. shahii Type VI CRISPR-Cas system mediates the immune response by inhibiting translation through collateral tRNA cleavage.", publisher = "Belgrade : Institute of molecular genetics and genetic engineering", journal = "5th Belgrade Bioinformatics Conference", title = "Transcriptome-wide detection of RNA cleavage sites revealed tRNA cleavage by target-activated CRISPR-Cas13a effector", pages = "130-130", url = "https://hdl.handle.net/21.15107/rcub_imagine_2470" }
Kolesnik, M., Jain, I., Semenova, E.,& Severinov, K.. (2024). Transcriptome-wide detection of RNA cleavage sites revealed tRNA cleavage by target-activated CRISPR-Cas13a effector. in 5th Belgrade Bioinformatics Conference Belgrade : Institute of molecular genetics and genetic engineering., 130-130. https://hdl.handle.net/21.15107/rcub_imagine_2470
Kolesnik M, Jain I, Semenova E, Severinov K. Transcriptome-wide detection of RNA cleavage sites revealed tRNA cleavage by target-activated CRISPR-Cas13a effector. in 5th Belgrade Bioinformatics Conference. 2024;:130-130. https://hdl.handle.net/21.15107/rcub_imagine_2470 .
Kolesnik, Matvei, Jain, Ishita, Semenova, Ekaterina, Severinov, Konstantin, "Transcriptome-wide detection of RNA cleavage sites revealed tRNA cleavage by target-activated CRISPR-Cas13a effector" in 5th Belgrade Bioinformatics Conference (2024):130-130, https://hdl.handle.net/21.15107/rcub_imagine_2470 .