grna coding sequence  (Addgene inc)

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Effects of PAM sites on editing efficiency of MAD7 construct 8 at the GUT1 locus.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: Construct, Clone Assay, Disruption

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: The main features of CRISPR-Cas9 and CRISPR-MAD7 systems.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: CRISPR

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Target genes for setting up the CRISPR-MAD7 system.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: CRISPR, Sequencing, Fluorescence

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Schematic representation of the CRISPR-MAD7 ( a ) and CRISPR-Cas9 ( b ) E. coli / K. phaffii shuttle vectors. Vector ( a ) encodes the gene for the expression of MAD7 (blue), whereas ( b ) encodes the genes for the expression of Cas9 (red). In both cases, the transcription is initiated by one side of the bidirectional, constitutive RNA polymerase II promotor HTX1 (P HTX1 ) and terminated by the DAS1 terminator ( DAS1 TT). Transcription of the sgRNA cassette (green) is initiated by the second side of the P HTX1 and terminated by the AOX1 terminator ( AOX1 TT). For selection purposes in E. coli and K. phaffii , a Zeocin resistance gene (Sh ble, purple) under the control of P ILV5 and P EM72 is used. For transient expression of the CRISPR elements in K. phaffii , the plasmids harbor an autonomously replicating sequence (PARS1, grey). For plasmid propagation during cloning procedures in E. coli , an origin of replication (pUC Ori, yellow) is encoded.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: CRISPR, Plasmid Preparation, Expressing, Selection, Sequencing, Clone Assay

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Screening plate for GUT1 deficient K. phaffii clones. After transformation with the CRISPR-MAD7 vectors, K. phaffi transformants were cultivated in 96-deep-well plates containing 250 µL YPD media for 48 h at 28 °C and 320 pm. Cell material from the cultivations was transferred onto screening plates with buffered minimal media plates containing 1.34% YNB, 4 × 10 −5 % biotin, 200 mM potassium phosphate buffer (pH 6.0), and 1% glycerol as carbon source using a metallic stamp. After 48 h at 28 °C, the number of mutants with a reduced growth phenotype on the glycerol media was evaluated: Clones in box A show wildtype-like growth (with the exception of one spot that was marked with an x to be excluded from this visualization); box B represents mutants with reduced growth, which are regarded as GUT1 deficient in this experiment.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: Clone Assay, Transformation Assay, CRISPR

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Editing efficiencies of MAD7 and sgRNA constructs at the GUT1 locus.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: Construct, CRISPR, Clone Assay, Disruption

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Effects of PAM sites on editing efficiency of MAD7 construct 8 at the GUT1 locus.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: Construct, Clone Assay, Disruption

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Disruption efficiency of CRISPR-MAD7 construct 21 targeting DsRed and Sh ble.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: Disruption, CRISPR, Construct, Clone Assay

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Summary of editing results using Cas9 and MAD7 endonucleases.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques:

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Summary of preferable PAM sites for the CRISPR-MAD7 system.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: CRISPR

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Summary of preferable PAM sites for the CRISPR-Cas9 system.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: CRISPR

Journal: Journal of Fungi

Article Title: Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii

doi: 10.3390/jof10030197

Figure Lengend Snippet: Summary of HDR-mediated editing efficiency.

Article Snippet: Moreover, 44 CRISPR-MAD7 plasmids with specific gRNA coding sequences were ordered from Twist Bioscience (San Francisco, CA, USA).

Techniques: