Monash Functional Genomics Platform

The cloning of a single guide RNA into a vector of either PLKO, Pxpr003, CROP SEQ guide puro (with or without EGFP/RFPTurbo) or Pxpr_053.

The cloning of a single guide RNA into a vector of either PLKO, Pxpr003, CROP SEQ guide puro (with or without EGFP/RFPTurbo) or Pxpr_053. This is for up to 5x sgRNAs. 

This is a pooled screen for adherent cell lines under low attachment conditions for up to 4 replicates with either a provided library DNA or the standard genome wide library (human Brunello OR mouse Brie). Cas9-P2A-Blast lentivirus is provided and Prpx011-GFP-Puro lentivirus is included to validate the activate of the Cas9 infection. This includes a titration and mycoplasma test of the provided cell line followed by performing the pooled screen on ultra-low attachment plates to provide the added challenge to proliferation and assess colony formation. The screen is prepared in house for sequencing. Sequencing results are analysed and interpreted by our bioinformatician to provide feedback and potential future directions of the project. 

This is a pooled screen for up to 4 replicates with either a provided library DNA or the standard genome wide library (human Brunello OR mouse Brie). Cas9-P2A-Blast lentivirus is provided and Prpx011-GFP-Puro lentivirus is included to validate the activate of the Cas9 infection. This includes a titration and mycoplasma test of the provided cell line followed by performing the pooled screen. The screen is prepared in house for sequencing. Sequencing results are analysed and interpreted by our bioinformatician to provide feedback and potential future directions of the project. 

This is a pooled screen for up to 4 replicates with either a provided library DNA or the standard genome wide library (human Brunello OR mouse Brie). Cas9-P2A-Blast lentivirus is provided and Prpx011-GFP-Puro lentivirus is included to validate the activate of the Cas9 infection. This includes a titration and mycoplasma test of the provided cell line followed by performing the pooled screen. The screen is prepared in house for sequencing. Sequencing results are analysed and interpreted by our bioinformatician to provide feedback and potential future directions of the project. 

Discuss with us about the details of the customised pooled screen and we can tailor the experiment to your needs while providing the best possible price. 

This is a pooled screen for up to 8 replicates with either a provided library DNA or the standard genome wide library (human Brunello OR mouse Brie). Cas9-P2A-Blast lentivirus is provided and Prpx011-GFP-Puro lentivirus is included to validate the activate of the Cas9 infection. This includes a titration and mycoplasma test of the provided cell line followed by performing the pooled screen. The screen is prepared in house for sequencing. Sequencing results are analysed and interpreted by our bioinformatician to provide feedback and potential future directions of the project. 

CROP Seq is a cutting edge technology that allows for single celled RNA seq of a library or around 10 000 cells per experiment. This project is financially assisted with the help of Phenomics Australia covering approximately 50% of the cost of the project. The price provided is the remaining share to be covered by the user. 

This includes the generation of the pooled library DNA for a CROP Seq (Single Celled RNA Seq of a pooled screen) pooled screen, the generation of lentivirus for the pooled screen and the pooled screen itself. 

Production of ≥ 100 ug of pDNA of a pooled library. This includes the library preparation and confirmation of the guide RNAs via next gen pooled sequencing for libraries for 12 000 to 92 000 guides.

Production of ≥ 100 ug of pDNA of a pooled library. This includes the library preparation and confirmation of the guide RNAs via next gen pooled sequencing for libraries up to 12 000 guides. 

We provide library preparation for next generation sequencing. This service allows you to provide genomic DNA from the screen, we then prepare it and provide you the the bioformatic results and intepretation of the data. The number of replicates is dependent upon the size of the library. Up to 15 libraries/replicates for those under 10 000 guides & up to 6 libraries/replicates for libraries of 70 000 guides. 

The Monash Functional Genomics Platform offers comprehensive advice on project/hypothesises design and grant proposals. Mechanistic characterisation can be devised using High-throughput expression profiling Data can be analysed through our integrated Bioinformatics capabilities, providing a rapid and unbiased approach to define the mechanisms associated with altered gene function in different Genome-wide or custom genetic screens biological models. Assistance can be provided for the generation of suitable Cas9 expressing cells and screens performed with gain and/or loss of function libraries.

These libraries can be genome-wide or smaller custom-designed gene sets specific for your research area. These approaches enable unbiased identification of new genes that contribute to cellular activities. The data generated will further understanding of the genetic networks associated with biological processes / disease and, for example, can assist with drug development or elucidate mechanisms of physiological phenomena.

Contact us for further information.

Pooled screening is faster, less labour-intensive and more cost- effective than array-based screens, making functional genomics more accessible for researchers investigating questions ranging from fundamental biology to addressing clinical needs. The Functional Genomics Platform provides the tools and expertise for unbiased, systematic, high- throughput gain-of-function and loss-of-function screens in human and mouse cells.

In collaboration with the MTRP network, we deliver a complete gene discovery and characterisation service. Our pooled library screens take advantage of the latest in CRISPRCas9 technology (CRISPRko,  CRISPRi, CRISPRa) based editors, single-cell sequencing and ORF libraries. We have validated genome-wide libraries available or we can work with you to custom-design libraries that suit your research needs.

Our team has a wealth of expertise in genomics, pooled library screening and life science research, and is dedicated to delivering excellent science across all disciplines. We encourage researchers to meet with us before starting an experiment. We can provide ideas and insight into the most effective pooled screen to use and how to get maximum value from the data generated. Our facility works closely with  Micromon, FlowCore, Genome Modification and Bioinformatics to provide a complete package of support from project design through to analysis, secondary screening and mechanistic characterization of gene targets.

Specialist Services

• Crop-Seq (Human and Mouse) Pooled screen for single-cell mRNA profiling after gene perturbation (200-1,000 perturbations)
• CRISPRko (CRISPR knockout) whole-genome  human and mouse libraries containing unique gene targeting sgRNAs plus controls for mediating gene knockout.
• CRISPRi (CRISPR interference) whole-genome  human and mouse libraries containing unique gene targeting sgRNAs plus controls for mediating suppression of gene expression.
• CRISPRa (CRISPR activation) whole-genome  human and mouse libraries containing unique gene targeting sgRNAs plus controls for activating gene expression.
• ORF (human) Single library containing 16,000 human protein-coding sequences. This library contains unique barcoded ORFs enabling pooled ORF screens.
• Custom CRISPR/Cas9 libraries can be designed to meet specific research needs and are unique to each project.
• CRISPR/Cas9 Cell line generation
• Bioinformatics analysis
  
  

Working with us 

• Fee for service
• Consultancies

Production of 9ml of a replication deficient lentivirus in DMEM 30% FBS at standard particle concentration from provided DNA for sgRNAs or other provided vectors. 

Production of 9ml of a replication deficient lentivirus designed to induce double stranded breaks in safe harbour genes. This acts as effective negative controls in experiments when following up hits from pooled screens. 

AAVS1 and ROSA26 guides are available in Pxpr003 or CROPseq-puro mCherry/GFP vectors. A choice from 5 different AAVS1 guides and 2 different mROSA26 guides are available.

Production of 9ml of a replication deficient lentivirus in DMEM 30% FBS at standard particle concentration from provided DNA for up to 5 sgRNAs or other provided vectors. 

This includes 5ml of the desired lentivirus for CRISPRc/i/a vectors with blasticidin resistance. 

This includes 5ml of Pxpr011 lentivirus to validate the effectiveness of Cas9 infection. 

The Monash Functional Genomics Platform offers comprehensive advice on project/hypothesises design and grant proposals. Mechanistic characterisation can be devised using High-throughput expression profiling Data can be analysed through our integrated Bioinformatics capabilities, providing a rapid and unbiased approach to define the mechanisms associated with altered gene function in different Genome-wide or custom genetic screens biological models. Assistance can be provided for the generation of suitable Cas9 expressing cells and screens performed with gain and/or loss of function libraries.

These libraries can be genome-wide or smaller custom-designed gene sets specific for your research area. These approaches enable unbiased identification of new genes that contribute to cellular activities. The data generated will further understanding of the genetic networks associated with biological processes / disease and, for example, can assist with drug development or elucidate mechanisms of physiological phenomena.

Contact us for further information.

Pooled screening is faster, less labour-intensive and more cost- effective than array-based screens, making functional genomics more accessible for researchers investigating questions ranging from fundamental biology to addressing clinical needs. The Functional Genomics Platform provides the tools and expertise for unbiased, systematic, high- throughput gain-of-function and loss-of-function screens in human and mouse cells.

In collaboration with the MTRP network, we deliver a complete gene discovery and characterisation service. Our pooled library screens take advantage of the latest in CRISPRCas9 technology (CRISPRko,  CRISPRi, CRISPRa) based editors, single-cell sequencing and ORF libraries. We have validated genome-wide libraries available or we can work with you to custom-design libraries that suit your research needs.

Our team has a wealth of expertise in genomics, pooled library screening and life science research, and is dedicated to delivering excellent science across all disciplines. We encourage researchers to meet with us before starting an experiment. We can provide ideas and insight into the most effective pooled screen to use and how to get maximum value from the data generated. Our facility works closely with  Micromon, FlowCore, Genome Modification and Bioinformatics to provide a complete package of support from project design through to analysis, secondary screening and mechanistic characterization of gene targets.

Specialist Services

• Crop-Seq (Human and Mouse) Pooled screen for single-cell mRNA profiling after gene perturbation (200-1,000 perturbations)
• CRISPRko (CRISPR knockout) whole-genome  human and mouse libraries containing unique gene targeting sgRNAs plus controls for mediating gene knockout.
• CRISPRi (CRISPR interference) whole-genome  human and mouse libraries containing unique gene targeting sgRNAs plus controls for mediating suppression of gene expression.
• CRISPRa (CRISPR activation) whole-genome  human and mouse libraries containing unique gene targeting sgRNAs plus controls for activating gene expression.
• ORF (human) Single library containing 16,000 human protein-coding sequences. This library contains unique barcoded ORFs enabling pooled ORF screens.
• Custom CRISPR/Cas9 libraries can be designed to meet specific research needs and are unique to each project.
• CRISPR/Cas9 Cell line generation
• Bioinformatics analysis
  
  

Working with us 

• Fee for service
• Consultancies

This is a pooled screen for up to 4 replicates with either a provided library DNA or the standard genome wide library (human Brunello OR mouse Brie). Cas9-P2A-Blast lentivirus is provided and Prpx011-GFP-Puro lentivirus is included to validate the activate of the Cas9 infection. This includes a titration and mycoplasma test of the provided cell line followed by performing the pooled screen. The screen is prepared in house for sequencing. Sequencing results are analysed and interpreted by our bioinformatician to provide feedback and potential future directions of the project. 

This is a pooled screen for up to 8 replicates with either a provided library DNA or the standard genome wide library (human Brunello OR mouse Brie). Cas9-P2A-Blast lentivirus is provided and Prpx011-GFP-Puro lentivirus is included to validate the activate of the Cas9 infection. This includes a titration and mycoplasma test of the provided cell line followed by performing the pooled screen. The screen is prepared in house for sequencing. Sequencing results are analysed and interpreted by our bioinformatician to provide feedback and potential future directions of the project. 

This is a pooled screen for adherent cell lines under low attachment conditions for up to 4 replicates with either a provided library DNA or the standard genome wide library (human Brunello OR mouse Brie). Cas9-P2A-Blast lentivirus is provided and Prpx011-GFP-Puro lentivirus is included to validate the activate of the Cas9 infection. This includes a titration and mycoplasma test of the provided cell line followed by performing the pooled screen on ultra-low attachment plates to provide the added challenge to proliferation and assess colony formation. The screen is prepared in house for sequencing. Sequencing results are analysed and interpreted by our bioinformatician to provide feedback and potential future directions of the project. 

This is a pooled screen for up to 4 replicates with either a provided library DNA or the standard genome wide library (human Brunello OR mouse Brie). Cas9-P2A-Blast lentivirus is provided and Prpx011-GFP-Puro lentivirus is included to validate the activate of the Cas9 infection. This includes a titration and mycoplasma test of the provided cell line followed by performing the pooled screen. The screen is prepared in house for sequencing. Sequencing results are analysed and interpreted by our bioinformatician to provide feedback and potential future directions of the project. 

Discuss with us about the details of the customised pooled screen and we can tailor the experiment to your needs while providing the best possible price. 

Production of ≥ 100 ug of pDNA of a pooled library. This includes the library preparation and confirmation of the guide RNAs via next gen pooled sequencing for libraries up to 12 000 guides. 

Production of ≥ 100 ug of pDNA of a pooled library. This includes the library preparation and confirmation of the guide RNAs via next gen pooled sequencing for libraries for 12 000 to 92 000 guides.

We provide library preparation for next generation sequencing. This service allows you to provide genomic DNA from the screen, we then prepare it and provide you the the bioformatic results and intepretation of the data. The number of replicates is dependent upon the size of the library. Up to 15 libraries/replicates for those under 10 000 guides & up to 6 libraries/replicates for libraries of 70 000 guides. 

CROP Seq is a cutting edge technology that allows for single celled RNA seq of a library or around 10 000 cells per experiment. This project is financially assisted with the help of Phenomics Australia covering approximately 50% of the cost of the project. The price provided is the remaining share to be covered by the user. 

This includes the generation of the pooled library DNA for a CROP Seq (Single Celled RNA Seq of a pooled screen) pooled screen, the generation of lentivirus for the pooled screen and the pooled screen itself. 

The cloning of a single guide RNA into a vector of either PLKO, Pxpr003, CROP SEQ guide puro (with or without EGFP/RFPTurbo) or Pxpr_053.

The cloning of a single guide RNA into a vector of either PLKO, Pxpr003, CROP SEQ guide puro (with or without EGFP/RFPTurbo) or Pxpr_053. This is for up to 5x sgRNAs. 

Production of 9ml of a replication deficient lentivirus in DMEM 30% FBS at standard particle concentration from provided DNA for sgRNAs or other provided vectors. 

Production of 9ml of a replication deficient lentivirus in DMEM 30% FBS at standard particle concentration from provided DNA for up to 5 sgRNAs or other provided vectors. 

This includes 5ml of the desired lentivirus for CRISPRc/i/a vectors with blasticidin resistance. 

This includes 5ml of Pxpr011 lentivirus to validate the effectiveness of Cas9 infection. 

Production of 9ml of a replication deficient lentivirus designed to induce double stranded breaks in safe harbour genes. This acts as effective negative controls in experiments when following up hits from pooled screens. 

AAVS1 and ROSA26 guides are available in Pxpr003 or CROPseq-puro mCherry/GFP vectors. A choice from 5 different AAVS1 guides and 2 different mROSA26 guides are available.