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Endogenous tagging using split mNeonGreen in human iPSCs for live imaging studies

Author(s): Husser MC; Pham NP; Law C; Araujo FRB; Martin VJJ; Piekny A;

Endogenous tags have become invaluable tools to visualize and study native proteins in live cells. However, generating human cell lines carrying endogenous tags is difficult due to the low efficiency of homology-directed repair. Recently, an engineered split mNeonGreen protein was used to generate a large-scale endogenous tag library in HEK293 cells. Usin ...

Article GUID: 38652106


CRISPR/Cas9 mediated gene editing of transcription factor ACE1 for enhanced cellulase production in thermophilic fungus Rasamsonia emersonii

Author(s): Singh V; Raheja Y; Basotra N; Sharma G; Tsang A; Chadha BS;

Background: The filamentous fungus Rasamsonia emersonii has immense potential to produce biorefinery relevant thermostable cellulase and hemicellulase enzymes using lignocellulosic biomass. Previously in our lab, a hyper-cellulase producing strain of R. emersonii was developed through classical breeding and system biology approaches. ACE1, a pivotal trans ...

Article GUID: 37658430


CRAPS: Chromosomal-Repair-Assisted Pathway Shuffling in Yeast

Author(s): Dykstra CB; Pyne ME; Martin VJJ;

A fundamental challenge of metabolic engineering involves assembling and screening vast combinations of orthologous enzymes across a multistep biochemical pathway. Current pathway assembly workflows involve combining genetic parts ex vivo and assembling one pathway configuration per tube or well. Here, we present CRAPS, Chromosomal-Repair-Assisted Pathway ...

Article GUID: 37584634


Rapid, scalable, combinatorial genome engineering by marker-less enrichment and recombination of genetically engineered loci in yeast

Author(s): Abdullah M; Greco BM; Laurent JM; Garge RK; Boutz DR; Vandeloo M; Marcotte EM; Kachroo AH;

A major challenge to rationally building multi-gene processes in yeast arises due to the combinatorics of combining all of the individual edits into the same strain. Here, we present a precise and multi-site genome editing approach that combines all edits without selection markers using CRISPR-Ca ...

Article GUID: 37323580


Cytokinetic diversity in mammalian cells is revealed by the characterization of endogenous anillin, Ect2 and RhoA

Author(s): Husser MC; Ozugergin I; Resta T; Martin VJJ; Piekny AJ;

Cytokinesis is required to physically separate the daughter cells at the end of mitosis. This crucial process requires the assembly and ingression of an actomyosin ring, which must occur with high fidelity to avoid aneuploidy and cell fate changes. Most of our knowledge of mammalian cytokinesis was generated using over-expressed transgenes in HeLa cells. ...

Article GUID: 36416720


The MyLo CRISPR-Cas9 Toolkit: A Markerless Yeast Localization and Overexpression CRISPR-Cas9 Toolkit

Author(s): Bean BDM; Whiteway M; Martin VJJ;

The genetic tractability of the yeast Saccharomyces cerevisiae has made it a key model organism for basic research and a target for metabolic engineering. To streamline the introduction of tagged genes and compartmental markers with powerful CRISPR-Cas9-based genome editing tools we constructed a Markerless Yeast Localization and Overexpression (MyLO) CRI ...

Article GUID: 35708612


The chimeric GaaR-XlnR transcription factor induces pectinolytic activities in the presence of D-xylose in Aspergillus niger

Author(s): Kun RS; Garrigues S; Di Falco M; Tsang A; de Vries RP;

Aspergillus niger is a filamentous fungus well known for its ability to produce a wide variety of pectinolytic enzymes, which have many applications in the industry. The transcriptional activator GaaR is induced by 2-keto-3-deoxy-L-galactonate, a compound derived from D-galacturonic acid, and plays a major role in the regulation of pectinolytic genes. The ...

Article GUID: 34236481


Identification of a Novel Biosynthetic Gene Cluster in Aspergillus niger Using Comparative Genomics

Author(s): Evdokias G; Semper C; Mora-Ochomogo M; Di Falco M; Nguyen TTM; Savchenko A; Tsang A; Benoit-Gelber I;

Previously, DNA microarrays analysis showed that, in co-culture with Bacillus subtilis, a biosynthetic gene cluster anchored with a nonribosomal peptides synthetase of Aspergillus niger is downregulated. Based on phylogenetic and synteny analyses, we show here that this gene cluster, NRRL3_00036- ...

Article GUID: 34064722


Using the endogenous CRISPR-Cas system of Heliobacterium modesticaldum to delete the photochemical reaction center core subunit gene.

Author(s): Baker PL, Orf GS, Kevershan K, Pyne ME, Bicer T, Redding KE

Appl Environ Microbiol. 2019 Sep 20;: Authors: Baker PL, Orf GS, Kevershan K, Pyne ME, Bicer T, Redding KE

Article GUID: 31540988


Single-step Precision Genome Editing in Yeast Using CRISPR-Cas9.

Author(s): Akhmetov A, Laurent JM, Gollihar J, Gardner EC, Garge RK, Ellington AD, Kachroo AH, Marcotte EM

Bio Protoc. 2018 Mar 20;8(6): Authors: Akhmetov A, Laurent JM, Gollihar J, Gardner EC, Garge RK, Ellington AD, Kachroo AH, Marcotte EM

Article GUID: 29770349


A Highly Characterized Synthetic Landing Pad System for Precise Multicopy Gene Integration in Yeast.

Author(s): Bourgeois L, Pyne ME, Martin VJJ

ACS Synth Biol. 2018 Nov 16;7(11):2675-2685 Authors: Bourgeois L, Pyne ME, Martin VJJ

Article GUID: 30372609


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