Lysate-based Systems

Lysate-based systems are cell-free platforms that use disrupted cells to retain biological machinery outside an intact living organism, enabling controlled biochemical reactions for bioengineering. Cell lysis releases enzymes, ribosomes, nucleic acids, and other molecular components into an extract, where supplied DNA, RNA, substrates, or cofactors can drive processes such as transcription, translation, and metabolic conversion. By separating these functions from cellular growth and survival, lysate-based systems support rapid prototyping of genetic circuits, biosensors, and engineered metabolic pathways. They also provide tractable models for studying molecular mechanisms and can advance distributed biomanufacturing and other applications requiring flexible, programmable biological production.

Lysate-based Systems - Related Videos

Research

JoVE Journal - Bioengineering
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Liquid Chromatography Coupled to Refractive Index or Mass Spectrometric Detection for Metabolite Profiling in Lysate-based Cell-free Systems

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Cited by 6 •

2021

The protocols describe high-performance liquid chromatography methods coupled to refractive index or mass spectrometric detection for studying metabolic reactions in complex lysate-based cell-free systems.

Research

JoVE Journal - Biology

HeLa Based Cell Free Expression Systems for Expression of Plasmodium Rhoptry Proteins

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Cited by 16 •

2015

Expression of malarial proteins in cell based systems remains challenging. We demonstrate two step and one step IVT (in vitro translation) cell free expression systems for expressing malarial recombinant rhoptry proteins from HeLa cells. We use a Ni-resin affinity based purification system to purify the rhoptry proteins.

Anion-Exchange Chromatography-Based Protein Purification: A Separation Technique to Isolate a Protein of Interest From Dialyzed Bacterial Lysate Based on Net Charge

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2025

In this video, we demonstrate an anion-exchange chromatography technique to purify human calcium-binding protein S100A12 based on the net surface charge of the protein from an Escherichia coli culture.

Peptide Purification: An RP-HPLC-based Technique to Extract Peptides from Digested Protein Lysates

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2023

In this video, we demonstrate reversed-phase high-performance liquid chromatography to purify peptides from a digested protein lysate mixture. These purified peptides are then lyophilized and stored for downstream applications. Analysis of peptides can help understand the mechanics involved in cellular signaling and cancer.

Preparation of Bacteriophage Lysate for Generalized Transduction

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2026

Source: Saragliadis, A., et al. Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes. J. Vis. Exp.(2018)This video demonstrates preparation of a bacteriophage lysate containing transducing particles. A donor strain is infected, plaques form, and the soft agar overlay is processed with chloroform to yield a clarified lysate for downstream analysis.

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