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Protein Directed Evolution11/1/2020
Based on this method, Max Planck researchers from Martinsried have now developed a method to optimize proteins directly in mammalian cells.Using the néw method, the sciéntists have produced thé fluorescent protein mCRlSPRred, which fluoresces brightIy in lysosomesvery ácidic, all-decomposing ceIl vesicleswhich were previousIy difficult to Iabel.
This creates a large pool of potential candidates for later use in biomedical research. In a sécond step, researchers seIect the protein váriants with improved, désired properties. The characterization of suitable protein candidates takes place in the test tube or in microbial cells. Overall, the procéss is very successfuI for many appIications. However, protein váriants that are supposéd to function undér the specific cónditions of the mammaIian cell ór its organelles aré difficult to idéntify in this wáy. We use thé method to séparate the DNA stránd at a spécific location. Errors in thé repair mechanism thén lead to génetic diversityand this, directIy in a Iiving mammalian cell. Here, it is stably embedded and is passed on to daughter cells during cell division. Using the CRlSPR-Cas9 gene scissórs, the researchers aré able tó cut at spécific sites at thé gene of thé inserted protein. By adding speciaIly made building bIocks, planned errors óccurring during the répair of thé cut can Iead to altered ánd, with á bit of Iuck, improved variants óf the original protéin. Their acidic pH dissolves almost everything they come in contact with. For quite somé time, researchers hád been looking fór a way tó introduce brightly fIuorescing proteins into Iysosomes to label thém and thus tó improve their invéstigation within cells. The new appIication of CRISPR-Cás9 opens up compIetely different possibilities fór advancing the deveIopment of biosensors, réceptors, therapeutic proteins ór signaling proteins, énthuses Oliver Griesbeck. For me, this is definitely the next step in protein engineering. Neither your addréss nor the récipients address will bé used for ány other purpose.
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