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High Throughput CRISPR/Cas9 cell line generation using the CellRaft Array

Award Number: R44GM134993
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 05/01/2017
PERIOD OF PERFORMANCE END DATE: 04/30/2022

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High Throughput CRISPR/Cas9 cell line generation using the CellRaft Array - Project Summary Genome editing technologies, such as CRISPR/Cas9 provide a rapid, and targeted means of both knocking out gene expression and knocking in gene modifications. Since our initial Phase I submission, the utility of CRISPR technology has expanded beyond the generation of cell lines, to forward genetic screening, in vivo manipulation of gene expression and even human therapeutics. Phase I efforts successfully demonstrated the use of the CellRaft Technology in a streamlined workflow for CRISPR-mediated genome editing in cell lines. Our Phase I report demonstrates several unique capabilities of the CellRaft technology for establishing genome edited cell lines using CRISPR: 1) performing all workflow steps (transfection, sorting, colony growth) on a single cell culture consumable; 2) releasing colonies from the array without disturbing the growth of other colonies (i.e. individual colony isolation, as opposed to en masse colony collection via trypsin); 3) sorting cells and colonies via imaging without requiring flow-based sorting methods which can damage cell health and perturb native phenotypes. By fully integrating the CRISPR workflow on a single platform, the CellRaft Array and the automated CellRaft AIR™ System, the genome editing process will be dramatically streamlined. During Phase II we will continue validating this workflow and prepare for commercialization on the broader genome editing market. We will both scale up manufacturing of high-throughput CellRaft Arrays tailored to CRISPR/Cas9 genome editing under multiple conditions at once, as well as validate the performance of the system at two external laboratories. A new software package is also proposed which enables investigators to track transfection positive cells during their initial clonal colony growth phase. This software platform will also allow users to track the growth of colonies emerging from transfection positive single cells and sort them based on temporal propagation characteristics. Subawardee William Marzluff, PhD of UNC-Chapel Hill will evaluate the new high-throughput CellRaft Arrays as well as the new colony tracking software package and evaluate the AIR™ System has a multi-lab core facility instrument. In a second subaward program, Mike McConnell, PhD of the University of Virginia will perform a time-course CRISPR experiment using the CellRaft Technology and automated AIR™ System. Using both CRISPR-mediated genome editing and time-course tracking of colony growth, he will develop an in vitro model of tuberous sclerosis by editing the TSC1 gene in human IPSCs. Based on discussions with several CellRaft customers who use the system for CRISPR-based assays, there remains a clearly unmet need for a platform which broadly supports CRISPR genome editing workflows. The CellRaft Technology’s combination of imaging capabilities, support for the culture of viable cells and colonies and ability to sort and isolate cells for molecular analysis, lends itself to becoming a sufficiently flexible platform to enable a broad range of CRISPR-based experiments.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2022 ( Subtotal = $0 )
2022	2020	CELL MICROSYSTEMS INC	801 CAPITOLA DR	DURHAM	NC	27713	DURHAM	USA	Biomedical Research and Research Training	000	3	8/28/2022	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2020 ( Subtotal = $819,896 )
2020	2020	CELL MICROSYSTEMS, INC.	801 CAPITOLA DR STE 10	DURHAM	NC	27713	DURHAM	USA	Biomedical Research and Research Training	001	3	4/17/2020	NON-COMPETING CONTINUATION	$819,896
2020	2019	CELL MICROSYSTEMS, INC.	801 CAPITOLA DR STE 10	DURHAM	NC	27713	DURHAM	USA	Biomedical Research and Research Training	000	2	1/16/2020	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$0
														Subtotal = $819,896

Issue Date FY: 2019 ( Subtotal = $813,352 )
2019	2019	CELL MICROSYSTEMS, INC.	2 DAVIS DRIVE ROOM 119 AND 118	RESEARCH TRIANGLE PARK	NC	27709	DURHAM	USA	Biomedical Research and Research Training	000	2	4/8/2019	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$813,352
														Subtotal = $813,352

Grand Total All Awards = $1,633,248

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High Throughput CRISPR/Cas9 cell line generation using the CellRaft Array

Award Number: R44GM134993

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 05/01/2017

PERIOD OF PERFORMANCE END DATE: 04/30/2022

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