Team:DTU-Denmark/Part Collection

The PHEAST team presents our Part Collection:

CRISPR-Cas9 Toolbox for Engineering of Komagataella phaffii GS115

We developed this part collection to aid genetic engineering in K. phaffii GS115. We experimentally validated state of the art CRISPR based tools that enable the user easy and fast to genetically engineer K. phaffii GS115. The collection contains crRNAs and their associated homology for flanking the expression cassette of interest for chromosomal integration and/or deletion in K. phaffii GS115. The present collection comprises 58 parts for integrating genes in neutral integration sites in the four chromosomes of K. phaffii GS115 or deleting genes related to methanol to formaldehyde conversion. The parts can be combined in endless ways to engineer K. phaffii GS115 in the future.


crRNAs and Repair Templates for Deletion

Part name Part Type Description
BBa_K3841027 RNA gRNA for CRISPR mediated knockout of the aox1 gene in Komagataella phaffii here refered to as crRNA_GS115_AOX1_C1
BBa_K3841028 RNA gRNA for CRISPR mediated knockout of the aox1 gene in Komagataella phaffii here refered to as crRNA_GS115_AOX1_C2
BBa_K3841029 RNA gRNA for CRISPR mediated knockout of the aox2 gene in Komagataella phaffii here refered to as crRNA_GS115_AOX2_C1
BBa_K3841030 RNA gRNA for CRISPR mediated knockout of the aox2 gene in Komagataella phaffii here refered to as crRNA_GS115_AOX2_C2
BBa_K3841031 RNA gRNA for CRISPR mediated knockout of the adh2 gene in Komagataella phaffii here refered to as crRNA_GS115_ADH2_C1
BBa_K3841032 RNA gRNA for CRISPR mediated knockout of the adh2 gene in Komagataella phaffii here refered to as crRNA_GS115_ADH2_C2
BBa_K3841033 DNA Repair template for AOX1 knockout
BBa_K3841034 DNA Repair template for AOX2 knockout
BBa_K3841035 DNA Repair template for ADH2 knockout

crRNAs and Homology Arms for Integration

Part name Part Type Description
BBa_K3841016 RNA gRNA for CRISPR neutral insertion site 1 in Komagataella phaffii here refered to as gRNA1_Chr_1_Cand_1
BBa_K3841017 RNA gRNA for CRISPR neutral insertion site 2 in Komagataella phaffii here refered to as gRNA5_Chr_2_Cand_1
BBa_K3841018 RNA gRNA for CRISPR neutral insertion site 2 in Komagataella phaffii here refered to as gRNA6_Chr_2_Cand_2
BBa_K3841019 RNA gRNA for CRISPR neutral insertion site 3 in Komagataella phaffii here refered to as gRNA13_Chr_3_Cand_1
BBa_K3841020 RNA gRNA for CRISPR neutral insertion site 3 in Komagataella phaffii here refered to as gRNA14_Chr_3_Cand_2
BBa_K3841021 RNA gRNA for CRISPR neutral insertion site 4 in Komagataella phaffii here refered to as gRNA11_Chr_4_Cand_3
BBa_K3841078 RNA gRNA for CRISPR neutral insertion site 1 in Komagataella phaffii here refered to as gRNA2_Chr_1_Cand_2
BBa_K3841079 RNA gRNA for CRISPR neutral insertion site 1 in Komagataella phaffii here refered to as gRNA3_Chr_1_Cand_3
BBa_K3841080 RNA gRNA for CRISPR neutral insertion site 1 in Komagataella phaffii here refered to as gRNA4_Chr_1_Cand_4
BBa_K3841081 RNA gRNA for CRISPR neutral insertion site 2 in Komagataella phaffii here refered to as gRNA7_Chr_2_Cand_3
BBa_K3841082 RNA gRNA for CRISPR neutral insertion site 2 in Komagataella phaffii here refered to as gRNA8_Chr_2_Cand_4
BBa_K3841083 RNA gRNA for CRISPR neutral insertion site 3 in Komagataella phaffii here refered to as gRNA15_Chr_3_Cand_3
BBa_K3841084 RNA gRNA for CRISPR neutral insertion site 3 in Komagataella phaffii here refered to as gRNA16_Chr_3_Cand_4
BBa_K3841085 RNA gRNA for CRISPR neutral insertion site 4 in Komagataella phaffii here refered to as gRNA9_Chr_4_Cand_1
BBa_K3841086 RNA gRNA for CRISPR neutral insertion site 4 in Komagataella phaffii here refered to as gRNA10_Chr_4_Cand_2
BBa_K3841087 RNA gRNA for CRISPR neutral insertion site 4 in Komagataella phaffii here refered to as gRNA12_Chr_4_Cand_4
BBa_K3841037 DNA Homology arm for neutral insertion site 1 in Komagataella phaffii
BBa_K3841038 DNA Homology arm for neutral insertion site 1 in Komagataella phaffii
BBa_K3841039 DNA Homology arm for neutral insertion site 1 in Komagataella phaffii
BBa_K3841040 DNA Homology arm for neutral insertion site 1 in Komagataella phaffii
BBa_K3841041 DNA Homology arm for neutral insertion site 1 in Komagataella phaffii
BBa_K3841042 DNA Homology arm for neutral insertion site 1 in Komagataella phaffii
BBa_K3841043 DNA Homology arm for neutral insertion site 1 in Komagataella phaffii
BBa_K3841044 DNA Homology arm for neutral insertion site 1 in Komagataella phaffii
BBa_K3841045 DNA Homology arm for neutral insertion site 2 in Komagataella phaffii
BBa_K3841046 DNA Homology arm for neutral insertion site 2 in Komagataella phaffii
BBa_K3841047 DNA Homology arm for neutral insertion site 2 in Komagataella phaffii
BBa_K3841048 DNA Homology arm for neutral insertion site 2 in Komagataella phaffii
BBa_K3841049 DNA Homology arm for neutral insertion site 2 in Komagataella phaffii
BBa_K3841050 DNA Homology arm for neutral insertion site 2 in Komagataella phaffii
BBa_K3841051 DNA Homology arm for neutral insertion site 2 in Komagataella phaffii
BBa_K3841052 DNA Homology arm for neutral insertion site 2 in Komagataella phaffii
BBa_K3841053 DNA Homology arm for neutral insertion site 3 in Komagataella phaffii
BBa_K3841054 DNA Homology arm for neutral insertion site 3 in Komagataella phaffii
BBa_K3841055 DNA Homology arm for neutral insertion site 3 in Komagataella phaffii
BBa_K3841056 DNA Homology arm for neutral insertion site 3 in Komagataella phaffii
BBa_K3841057 DNA Homology arm for neutral insertion site 3 in Komagataella phaffii
BBa_K3841058 DNA Homology arm for neutral insertion site 3 in Komagataella phaffii
BBa_K3841059 DNA Homology arm for neutral insertion site 3 in Komagataella phaffii
BBa_K3841060 DNA Homology arm for neutral insertion site 3 in Komagataella phaffii
BBa_K3841061 DNA Homology arm for neutral insertion site 4 in Komagataella phaffii
BBa_K3841062 DNA Homology arm for neutral insertion site 4 in Komagataella phaffii
BBa_K3841063 DNA Homology arm for neutral insertion site 4 in Komagataella phaffii
BBa_K3841064 DNA Homology arm for neutral insertion site 4 in Komagataella phaffii
BBa_K3841065 DNA Homology arm for neutral insertion site 4 in Komagataella phaffii
BBa_K3841066 DNA Homology arm for neutral insertion site 4 in Komagataella phaffii
BBa_K3841067 DNA Homology arm for neutral insertion site 4 in Komagataella phaffii
BBa_K3841068 DNA Homology arm for neutral insertion site 4 in Komagataella phaffii

gRNA Backbone

Part name Part Type Description
BBa_K3841003 DNA gRNA backbone used for Cas9 mediated double-stranded breaks.