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

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 9R44GM134993-02
Agency Tracking Number: R44GM134993
Amount: $813,352.00
Phase: Phase II
Program: SBIR
Solicitation Topic Code: 400
Solicitation Number: PA18-574
Solicitation Year: 2018
Award Year: 2019
Award Start Date (Proposal Award Date): 2019-05-01
Award End Date (Contract End Date): 2021-04-30
Small Business Information
907 GREENWOOD RD, Chapel Hill, NC, 27514-3912
DUNS: 962655853
HUBZone Owned: N
Woman Owned: N
Socially and Economically Disadvantaged: N
Principal Investigator
 Jessica Hartman
 (708) 218-8473
Business Contact
Phone: (708) 218-8473
Research Institution
Project SummaryGenome editing technologiessuch as CRISPR Casprovide a rapidand targeted means of both knocking out gene expression and knocking in gene modificationsSince our initial Phase I submissionthe utility of CRISPR technology has expanded beyond the generation of cell linesto forward genetic screeningin vivo manipulation of gene expression and even human therapeuticsPhase I efforts successfully demonstrated the use of the CellRaft Technology in a streamlined workflow for CRISPR mediated genome editing in cell linesOur Phase I report demonstrates several unique capabilities of the CellRaft technology for establishing genome edited cell lines using CRISPRperforming all workflow stepstransfectionsortingcolony growthon a single cell culture consumablereleasing colonies from the array without disturbing the growth of other coloniesi eindividual colony isolationas opposed to en masse colony collection via trypsinsorting cells and colonies via imaging without requiring flow based sorting methods which can damage cell health and perturb native phenotypesBy fully integrating the CRISPR workflow on a single platformthe CellRaft Array and the automated CellRaft AIRSystemthe genome editing process will be dramatically streamlinedDuring Phase II we will continue validating this workflow and prepare for commercialization on the broader genome editing marketWe will both scale up manufacturing of high throughput CellRaft Arrays tailored to CRISPR Casgenome editing under multiple conditions at onceas well as validate the performance of the system at two external laboratoriesA new software package is also proposed which enables investigators to track transfection positive cells during their initial clonal colony growth phaseThis 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 characteristicsSubawardee William MarzluffPhD of UNC Chapel Hill will evaluate the new high throughput CellRaft Arrays as well as the new colony tracking software package and evaluate the AIRSystem has a multi lab core facility instrumentIn a second subaward programMike McConnellPhD of the University of Virginia will perform a time course CRISPR experiment using the CellRaft Technology and automated AIRSystemUsing both CRISPR mediated genome editing and time course tracking of colony growthhe will develop an in vitro model of tuberous sclerosis by editing the TSCgene in human IPSCsBased on discussions with several CellRaft customers who use the system for CRISPR based assaysthere remains a clearly unmet need for a platform which broadly supports CRISPR genome editing workflowsThe CellRaft Technology s combination of imaging capabilitiessupport for the culture of viable cells and colonies and ability to sort and isolate cells for molecular analysislends itself to becoming a sufficiently flexible platform to enable a broad range of CRISPR based experiments Project NarrativeGenome editing by CRISPR Cashas become one of the more powerful molecular genetic methods available in contemporary researchThe method can be used to not only generate cell linesbut also to manipulate gene expression in vivo as well as screen thousands of genes in a single experiment for various phenotypes of interestOur Phase I efforts successfully demonstrated the utility of the CellRaft Technology to accelerate the development of CRISPR Casgenome edited cell linesIn this Phase II programwe will prepare for commercialization of this powerfulstreamlined workflowas well as expand the utility of the technology to CRISPR knockout disease models

* Information listed above is at the time of submission. *

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