Genome Editing Services

To place an order, please use the service request form. The HSCI iPS Core offers the following genome editing services (rates differ according to Harvard affiliation and profit/non-profit status):

Genome editing services
CRISPR technology modules Rates for Harvard and HSCI Faculty+ Rates for Non-profit Institutions Rates for For-profit institutions
CRISPR Module 1: Background study and gRNA design $474/CRISPR target $601/CRISPR target $1,016/CRISPR target
CRISPR Module 2a: CRISPR/Cas9-mediated mini knock-out $12,664/gene/hPSC line $15,826/gene/hPSC line $26,746/gene/hPSC line
CRISPR Module 2b: CRISPR/Cas9-mediated midi knock-out $14,575/gene/hPSC line $17,737/gene/hPSC line $29,976/gene/hPSC line
CRISPR Module 2c: CRISPR/Cas9-mediated maxi knock-out $16,594/hPSC line $19,756/hPSC line $33,387/hPSC line
CRISPR Module 3a: CRISPR/Cas9-mediated mini knock-in $14,575/gene/hPSC line $17,737/gene/hPSC line $29,976/gene/hPSC line
CRISPR Module 3b: CRISPR/Cas9-mediated midi knock-in $16,486/gene/hPSC line $19,648/gene/hPSC line $33,205/gene/hPSC line
CRISPR Module 3c: CRISPR/Cas9-mediated maxi knock-in $19,799/gene/hPSC line $22,961/gene/hPSC line $38,804/gene/hPSC line
CRISPR Module 4a: CRISPR/Cas9-mediated transgene insertion with permanent selection - one allele $12,720/insertion/hPSC line $15,912/insertion/hPSC line $26,892/insertion/hPSC line
CRISPR Module 4b: CRISPR/Cas9-mediated transgene insertion with excision of selection cassette - one allele $14,651/insertion/hPSC line $17,844/insertion/hPSC line $30,156/insertion/hPSC line
CRISPR Module 4c: CRISPR/Cas9-mediated transgene - other $14,631/insertion/hPSC line $17,823/insertion/hPSC line $30,122/insertion/hPSC line
CRISPR Module 5: transgene/selection cassette removal with Cre-loxP or PiggyBac $5,462/excision/hPSC line $6,284/excision/hPSC line $10,620/excision/hPSC line

+Note: "Harvard and HSCI Faculty" includes two groups of users: (1) All Harvard faculty and researchers using HU 33-digit charging code; and (2) faculty with HSCI appointments.

 

Process Guideline

CRISPR Module 1: Construction/Testing CRISPR vectors

This module, which takes about 2 weeks to complete, comprises consults with the investigator to discuss what the best CRISPR design and targeting strategy would be to generate an edited human pluripotent stem cell (hPSC) line with the investigators’ gene of interest targeted.

The module delivers customized construction of a CRISPR guide RNA plasmid containing a synthetic guide RNA complementary to the DNA sequence in the genomic site to be targeted.

Upon request the CRISPR guide RNA plasmid can be validated for nuclease efficiency by co-transfection of the CRISPR guide RNA plasmid and a Cas9-GFP plasmid using a genomic cleavage assay.

CRISPR Module 2: CRISPR/Cas9-mediated gene knock-out

This service, which takes about 6 months to complete, includes:

  • hPSC expansion/adaptation to single cell passaging
  • Mycoplasma testing
  • Electroporation of CRIPSR guide RNA and Cas9-GFP plasmids into 1 hPSC line
  • FACS sorting (pre-selection) of electroporated hPSCs
  • Clonal expansion of targeted line
  • PCR screening of targeted clones, followed by DNA Sanger sequencing of mutant clones to verify correct targeting
  • Expansion of mutant clones
  • Mycoplasma testing
  • Freezing (6 vials per line)

CRISPR Module 3: CRISPR/Cas9-mediated gene mutation introduction and/or repair

This service, which takes 6 to 7 months to complete, includes:

  • hPSC expansion/adaptation to single cell passaging
  • Mycoplasma testing
  • Electroporation of a CRISPR guide RNA, Cas9-GFP plasmid and a single stranded oligodinucleotide (ssODN) containing wildtype or mutated DNA sequence into 1 hPSC line
  • FACS sorting (pre-selection) electroporated hPSCs
  • Clonal expansion of targeted line
  • PCR/RE digest screening of targeted clones, followed by DNA Sanger sequencing to verify correct targeting and introduction of gene mutation/repair
  • Expansion of mutant clones
  • Mycoplasma testing
  • Freezing (6 vials per line)

For introducing or repairing mutations in hPSC lines in this module, we will design and have synthesized an ssODN containing the mutated or corrected DNA sequence, respectively.