Publications related to Molecular cloning

Development of cysteine cathepsin inhibitors combined with cell type-specific delivery for the treatment of B-cell lymphoma and solid tumors

Cysteine cathepsins proteases are enzymes that play essential physiological roles, but their activity is also associated to different aspects of cancer progression and to the development of other diseases. Therefore, cysteine cathepsins are relevant and pr ...

EPFL2024

Il-10 expressing cells for enhanced cancer immunotherapies

Li Tang, Yugang Guo, Yang Zhao

The present invention relates generally to the field of anti-cancer therapy, in particular to the use of adoptive T cell transfer therapy for treating cancer, in particular sold tumors. More specifically, the present invention relates to immune cells compr ...

2023

Design of an artificial phage-display library based on a new scaffold improved for average stability of the randomized proteins

Ghérici Hassaïne

Scaffold-based protein libraries are designed to be both diverse and rich in functional/folded proteins. However, introducing an extended diversity while preserving stability of the initial scaffold remains a challenge. Here we developed an original approa ...

Nature Portfolio2023

Microfluidic T Cell Selection by Cellular Avidity

Li Tang, Armand Kurum, Alexandre Harari

No T cell receptor (TCR) T cell therapies have obtained clinical approval. The lack of strategies capable of selecting and recovering potent T cell candidates may be a contributor to this. Existing protocols for selecting TCR T cell clones for cell therapi ...

WILEY2022

The enhancement of DNA fragmentation in a bench top ultrasonic water bath with needle-induced air bubbles: Simulation and experimental investigation

Martinus Gijs, Thomas Lehnert, Yang Liu, Lin Sun

Shearing DNA to a certain size is the first step in many medical and biological applications, especially in next-generation gene sequencing technology. In this article, we introduced a highly efficient ultrasonic DNA fragmentation method enhanced by needle ...

AIP Publishing2022

Considerations of the Impacts of Cell-Specific Growth and Production Rate on Clone Selection-A Simulation Study

Florian Maria Wurm

For the manufacturing of complex biopharmaceuticals using bioreactors with cultivated mammalian cells, high product concentration is an important objective. The phenotype of the cells in a reactor plays an important role. Are clonal cell populations showin ...

2021

A ribonucleoprotein transfection strategy for CRISPR/Cas9-mediated gene editing and single cell cloning in rainbow trout cells

Kristin Schirmer

Background The advent of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology marked the beginning of a new era in the field of molecular biology, allowing the efficient and precise creation of targeted mutations in the ge ...

2021

m6A methylation potentiates cytosolic dsDNA recognition in a sequence-specific manner

Andrea Ablasser, Muhammet Fatih Gülen

Nucleic acid sensing through pattern recognition receptors is critical for immune recognition of microbial infections. Microbial DNA is frequently methylated at the N-6 position of adenines (m6A), a modification that is rare in mammalian host DNA. We show ...

ROYAL SOC2021

Generation of a Large Peptide Phage Display Library by Self-Ligation of Whole-Plasmid PCR Product

Christian Heinis, Xudong Kong, Vanessa Carle, Cristina Diaz Perlas, Kaycie Marie Butler

The success of phage display, used for developing target-specific binders based on peptides and proteins, depends on the size and diversity of the library screened, but generating large libraries of phage-encoded polypeptides remains challenging. New pepti ...

2020

Traceable Impedance-Based Dispensing and Cloning of Living Single Cells

Philippe Renaud, Yann Barrandon, Ariane Rochat, David Vincent Bonzon, Stéphanie Claudinot, Georges Muller

Single-cell cloning is essential in stem cell biology, cancer research, and biotechnology. Regulatory agencies now require an indisputable proof of clonality that current technologies do not readily provide. Here, we report a one-step cloning method using ...

2020

Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology

Sebastian Maerkl, Laura Sophie Grasemann, Zoe Newell Swank, Barbora Lavickova, Grégoire Michielin, Nadanai Laohakunakorn, Amir Shahein

Cell-free systems offer a promising approach to engineer biology since their open nature allows for well-controlled and characterized reaction conditions. In this review, we discuss the history and recent developments in engineering recombinant and crude e ...

2020

Monobodies as enabling tools for structural and mechanistic biology

Oliver Hantschel

Monobodies, built with the scaffold of the fibronectin type III domain, are among the most well-established synthetic binding proteins. They promote crystallization of challenging molecular systems. They have strong tendency to bind to functional sites and ...

CURRENT BIOLOGY LTD2020

Molecular-Level Transformation of Dissolved Organic Matter during Oxidation by Ozone and Hydroxyl Radical

Urs von Gunten

Ozonation of drinking and wastewater relies on ozone (O-3) and hydroxyl radical ((OH)-O-center dot) as oxidants. Both oxidants react with dissolved organic matter (DOM) and alter its composition, but the selectivity of the two oxidants and mechanisms of re ...

AMER CHEMICAL SOC2020

Scalable Production of AAV Vectors in Orbitally Shaken HEK293 Cells

Florian Maria Wurm, Bernard Schneider, Vivianne Padrun, Nicole Déglon, Daniel Roman Blessing, Gabriel Vachey

Adeno-associated virus (AAV) vectors are currently among the most commonly applied for in vivo gene therapy approaches. The evaluation of vectors during clinical development requires the production of considerable amounts of highly pure and potent vectors. ...

2019

A new approach to Cas9-based genome editing in Aspergillus niger that is precise, efficient and selectable

Laure Marie Colette Leynaud-Kieffer Curran

Aspergillus niger and other filamentous fungi are widely used in industry, but efficient genetic engineering of these hosts remains nascent. For example, while molecular genetic tools have been developed, including CRISPR/Cas9, facile genome engineering of ...

2019

Bypassing bacterial infection in phage display by sequencing DNA released from phage particles

Christian Heinis, Xudong Kong, Camille Villequey

Phage display relies on a bacterial infection step in which the phage particles are replicated to perform multiple affinity selection rounds and to enable the identification of isolated clones by DNA sequencing. While this process is efficient for wild-typ ...

2017

Label-free identification of activated T lymphocytes through tridimensional microsensors on chip

Carlotta Guiducci, Elena Bianchi, George Coukos, Enrica Rollo, Enrico Tenaglia, Alexandre Harari

Label-free approaches to assess cell properties ideally suit the requirements of cell-based therapeutics, since they permit to characterize cells with minimal perturbation and manipulation, at the benefit of sample recovery and re-employment for treatment. ...

Elsevier2017

Recombinant protein production from stable mammalian cell lines and pools

David Hacker, Sowmya Balasubramanian

We highlight recent developments for the production of recombinant proteins from suspension-adapted mammalian cell lines. We discuss the generation of stable cell lines using transposons and lentivirus vectors (non-targeted transgene integration) and site- ...

Elsevier2016

Comparison of Three Transposons for the Generation of Highly Productive Recombinant CHO Cell Pools and Cell Lines

Florian Maria Wurm, David Hacker, Lucia Baldi Unser, Yashas Rajendra, Sowmya Balasubramanian

Several naturally occurring vertebrate transposable elements have been genetically modified to enable the transposition of recombinant genes in mammalian cells. We compared three transposonspiggyBac, Tol2, and Sleeping Beautyfor their ability to generate c ...

Wiley-Blackwell2016

Single Molecule Localization and Discrimination of DNA–Protein Complexes by Controlled Translocation Through Nanocapillaries

Aleksandra Radenovic, Sebastian Maerkl, Sanjin Marion, Roman Bulushev, Sebastian James Davis, Ekaterina Emilova Petrova

Through the use of optical tweezers we performed controlled translocations of DNA-protein complexes through nanocapillaries. We used RNA polymerase (RNAP) with two binding sites on a 7.2 kbp DNA fragment and a dCas9 protein tailored to have five binding si ...

American Chemical Society (ACS)2016