| 1. |
- Wimberger, Sandra, 1987, et al.
(författare)
-
Simultaneous inhibition of DNA-PK and Pol ϴ improves integration efficiency and precision of genome editing
- 2023
-
Ingår i: Nature Communications. ; 14:1
-
Tidskriftsartikel (refereegranskat)abstract
- Genome editing, specifically CRISPR/Cas9 technology, has revolutionized biomedical research and offers potential cures for genetic diseases. Despite rapid progress, low efficiency of targeted DNA integration and generation of unintended mutations represent major limitations for genome editing applications caused by the interplay with DNA double-strand break repair pathways. To address this, we conduct a large-scale compound library screen to identify targets for enhancing targeted genome insertions. Our study reveals DNA-dependent protein kinase (DNA-PK) as the most effective target to improve CRISPR/Cas9-mediated insertions, confirming previous findings. We extensively characterize AZD7648, a selective DNA-PK inhibitor, and find it to significantly enhance precise gene editing. We further improve integration efficiency and precision by inhibiting DNA polymerase theta (Pol ϴ). The combined treatment, named 2iHDR, boosts templated insertions to 80% efficiency with minimal unintended insertions and deletions. Notably, 2iHDR also reduces off-target effects of Cas9, greatly enhancing the fidelity and performance of CRISPR/Cas9 gene editing. Low efficiency of target DNA integration remains a challenge in genome engineering. Here the authors perform large-scale compound library and genetic screens to identify targets that enhance gene editing: they see that combined DNA-PK and Pol ϴ inhibition with potent compounds increases editing efficiency and precision.
|
|
| 2. |
- Demichev, Vadim, et al.
(författare)
-
A time-resolved proteomic and prognostic map of COVID-19
- 2021
-
Ingår i: Cell Systems. - : Elsevier BV. - 2405-4712 .- 2405-4720. ; 12:8, s. 780-794.e7
-
Tidskriftsartikel (refereegranskat)abstract
- COVID-19 is highly variable in its clinical presentation, ranging from asymptomatic infection to severe organ damage and death. We characterized the time-dependent progression of the disease in 139 COVID-19 inpatients by measuring 86 accredited diagnostic parameters, such as blood cell counts and enzyme activities, as well as untargeted plasma proteomes at 687 sampling points. We report an initial spike in a systemic inflammatory response, which is gradually alleviated and followed by a protein signature indicative of tissue repair, metabolic reconstitution, and immunomodulation. We identify prognostic marker signatures for devising risk-adapted treatment strategies and use machine learning to classify therapeutic needs. We show that the machine learning models based on the proteome are transferable to an independent cohort. Our study presents a map linking routinely used clinical diagnostic parameters to plasma proteomes and their dynamics in an infectious disease.
|
|
| 3. |
- Ekstrom, A., et al.
(författare)
-
Optimized Chiral Nucleon-Nucleon Interaction at Next-to-Next-to-Leading Order
- 2013
-
Ingår i: Physical Review Letters. - 1079-7114 .- 0031-9007. ; 110:19, s. Art. no. 192502-
-
Tidskriftsartikel (refereegranskat)abstract
- We optimize the nucleon-nucleon interaction from chiral effective field theory at next-to-next-to-leading order (NNLO). The resulting new chiral force NNLOopt yields chi(2) approximate to 1 per degree of freedom for laboratory energies below approximately 125 MeV. In the A = 3, 4 nucleon systems, the contributions of three-nucleon forces are smaller than for previous parametrizations of chiral interactions. We use NNLOopt to study properties of key nuclei and neutron matter, and we demonstrate that many aspects of nuclear structure can be understood in terms of this nucleon-nucleon interaction, without explicitly invoking three-nucleon forces.
|
|
