VIRAL VECTOR & GENE EDITING CORE
CRISPR/CAS9 Gene-Editing Tools
Overview
CRISPR ( C lustered R egularly I nterspaced S hort P alindromic R epeats) is a powerful tool that enables scientists to manipulate the genome with ease. Over the past decade, scientists have expanded the CRISPR/Cas arsenal to include many different types of genetic manipulations, including gene knock-outs, knock-ins, point mutations, base changes, and small and large insertions and deletions. For more information on the wide range of CRISPR technologies, please email us viral.core@med.fsu.edu
The advanced CRISPR/Cas tools will support your research as follows;
- Loss-of-function studies aimed to generate complete and permanent loss of gene expression or function (knock-out)
- Introduce a fragment-of-interest into a specific genomic locus (knock-in)
- Create a specific mutant allele of a gene (point mutant)
- Increase or decrease expression of a target gene (epigenome editing)
Vectors Available for CRISPR/CAS9 Expression
Lentiviral Vectors - backbones
- pBK109- short version derived from pLentiCRISPR/v2 ready-for-gRNA-cloning (BsmBI)
- pBK301- short version of pLentiCRISPR/v2 with 2 Sp1 binding sites upstream of U6p- gRNA cloning site BsmBI
- pBK175- short version of pLentiCRISPR/v2 with 4 Sp1 binding sites upstream of U6p- gRNA cloning site BsmBI
- pBK110- pLentiCRISPR/v2 Luciferase-gRNA
- pBK104- pLentiCRISPR/v2 GABA a2-subunit-gRNA
- pBK83- pLentiCRISPR/v2 HDAC1-1 –gRNA
- pBK84-pLentiCRISPR/v2 HDAC1-2 –gRNA
- pBK85-pLentiCRISPR/v2 HDAC1-3 – gRNA
- pBK86-pLentiCRISPR/v2 GFP1 – gRNA
- pBK87-pLentiCRISPR/v2 GFP2 – gRNA
- pBK88-pLentiCRISPR/v2 GFP3 – gRNA
- pBK189- pLentiCRISPR/v2 with GFP-gRNA
- pBK198- short version of pLentiCRISPR/v2 with 2 Sp1 binding sites with GFP-gRNA
- pBK180- rtTA3 Blasticidin third generation rtTA3 can be used for tet-Cas9 system (next plasmid)
- pBK185- tet- inducible Cas9- all-in-one short- with BsmBI for cloning sgRNA
- pBK195- tet- inducible Cas9- all-in-one short- with GFP-gRNA
- pCW-Cas9 Tet ON Plasmid #50661; Addgene- https://www.addgene.org/50661/
- pBK109easy -GFP pLenti-SpCas9gRNA with GFP all-in-one
- pBK97- SpCas9 nickase- all-in-one lentiviral backbone; titer-optimized backbone; ready for gRNA cloning BsmBI
- pBK456- dCas9- all-in-one lentiviral backbone; titer-optimized backbone; ready for gRNA cloning BsmBI
- pBK109easy -GFP pLenti-SpCas9gRNA with GFP all-in-one
- pBK109BL- short version of pLentiCRISPR/v2 gRNA cloning site BsmBI with blasticidin
- pBK114- pLenti-Cas9-GFP
- pLentiCRISPR/v2 Plasmid #52961; Addgene- https://www.addgene.org/52961/
- pLenti-multi-CRISPR; Plasmid #85402; Addgene- https://www.addgene.org/85402/
- LentiCRISPRv2-mCherry; Plasmid # 99154; Addgene- http://www.addgene.org/99154/
- pLenti-pU6-sgRNA Ef1alpha-Puro-T2A-BFP Plasmid #84832; Addgene- http://www.addgene.org/84832/
- pLenti-sgRNA; Plasmid #71409; Addgene- http://www.addgene.org/71409/
- pLenti-CRISPR.EFS.tRFP; Plasmid #57819; Addgene- http://www.addgene.org/57819/
- lentiCas9-Venus; Plasmid # 70267; Addgene- http://www.addgene.org/70267/
- pL-CRISPR.EFS.GFP Plasmid #57818; Addgene- https://www.addgene.org/57818/
- lentiCas9-Blast Plasmid #52962; Addgene- https://www.addgene.org/52962/
- lentiCas9-Puro Plasmid #52963; Addgene-http://www.addgene.org/52963/
- lentiCas9-EGFP; Plasmid #63592; Addgene- https://www.addgene.org/63592/
- lentiCas9n(D10A)-Blast; Plasmid #63593; Addgene- http://www.addgene.org/63593
- lenti-dCAS-VP64-Blast; Plasmid #61425; Addgene- http://www.addgene.org/61425/
- pHAGE TRE dCas9-VP64; Plasmid #50916; Addgene- http://www.addgene.org/50916/
- lenti-TRE-KRAB-dCas9-IRES-BFP; Plasmid #85449; Addgene- http://www.addgene.org/85449/
- plenti- hUbC-dCas9 VP64-T2A-GFP; Plasmid #53192; Addgene- http://www.addgene.org/53192/
- pKLV-U6gRNA(BbsI)-PGKpuro2ABFP; Plasmid #50946; Addgene- http://www.addgene.org/50946/
- plentiSAMv2; Plasmid #75112; Addgene- http://www.addgene.org/75112/
- pHR-SFFV-KRAB-dCas9-P2A-mCherry; Plasmid #60954; Addgene- http://www.addgene.org/60954/
- pLV-dCas9-KRAB-PGK-Hyg- Plasmid #83890; Addgene- http://www.addgene.org/83890/
- pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-GFP; Plasmid #71237; Addgene- http://www.addgene.org/71237/
- pHAGE EF1α dCas9-KRAB; Plasmid #50919; Addgene- http://www.addgene.org/50919/
- lenti sgRNA-MS2-Zeo; Plasmid #61427; Addgene- http://www.addgene.org/61427/
- lenti sgRNAMS2- puro optimized backbone; Plasmid #73797; Addgene- http://www.addgene.org/73797/
- pHAGE-TO-nmdCas9-3XGFP; Plasmid #64109; Addgene- http://www.addgene.org/64109/
- pLV-dCas9-p300-P2A-Puro; Plasmid #83889; Addgene- http://www.addgene.org/83889/
- Lenti_sgRNA-EFS-GFP; Plasmid #65656; Addgene- http://www.addgene.org/65656/
- Lenti-AsCpf1-Blast; Plasmid #84750; Addgene- http://www.addgene.org/84750/
- LentiCRISPRv2Cre; Plasmid #82415; Addgene- http://www.addgene.org/82415/
- Exp_v-pcDNA3.1-hAsCpf1; Plasmid #69982; Addgene- https://www.addgene.org/69982/
- Exp_v-pSimpleII-U6- gRNA cloning site BsmBI-NLS-NmCas9-HA-NLS plasmid # 47868; Addgene https://www.addgene.org/47868/
- Exp_v-pSaCas9_GFP; Plasmid # 64709; Addgene- https://www.addgene.org/64709/
- Exp_v-Csy4-T2A-Cas9-NLS; Plasmid # 53371; Addgene- https://www.addgene.org/53371/
- Exp_v-ppcDNA3.1-hFnCpf1; Plasmid #69976; Addgene- https://www.addgene.org/69976/
- Exp_v-eSpCas9(1.1)- Plasmid #71814; Addgene-https://www.addgene.org/71814/
- Exp_v-SpCas9-HF1 (high fidelity Cas9); Plasmid #72247; Addgene- https://www.addgene.org/72247/
AAVs
- AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA Plasmid # 61591; Addgene- https://www.addgene.org/61591/
- BK- pAAV-EFS-NC- SpCas9-NLS-Poly(A)
- BK- pAAV-CMV-SpCas9- NLS-Poly(A)
- BK- pAAV-hSyn -SpCas9- NLS-Poly(A)
- BK- pAAV- -SpCas9- NLS-Poly(A)
- pAAV- nEF promoter- Cas9; Plasmid #87115; Addgene- http://www.addgene.org/87115/
- pAAV-pMecp2-SpCas9 Plasmid #60957 http://www.addgene.org/60957/
- pX602-AAV-TBG::NLS-SaCas9-NLS-HA-OLLAS-bGHpA;U6::BsaI-sgRNA Plasmid #61593; Addgene- http://www.addgene.org/61593/
- pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA; Plasmid #61591; Addgene- http://www.addgene.org/61591/
- AAV-NFS-NC: NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA
- AAV-Syn: NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA
- pX600-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA; Plasmid #61592; Addgene- http://www.addgene.org/61592/
- pJEP12-AAV-H1/TO(dox-regulated)-L-sgRNA(Empty)-CMV-TetR-P2A-eGFP-KASH-pA; Plasmid #82704; Addgene- http://www.addgene.org/82704/
- pX603-AAV-CMV::NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA; Plasmid #61594; Addgene- http://www.addgene.org/61594/
- AAV-NFS-NC:NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA
- AAV-Syn:NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA
- AAV-MeCp2:NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA
- AAV-NFS-NC:NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA
- AAV-NFS-NC:NLS-dSaCas9Nickase-NLS-3xHA-bGHpA
- AAV-CMV:NLS-dSaCas9Nickase-NLS-3xHA-bGHpA
- AAV-CAG:NLS-dSaCas9Nickase NLS-3xHA-bGHpA
- AAV:ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR; Plasmid #60231; Addgene- http://www.addgene.org/60231/
- AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR; Plasmid #60226; Addgene- http://www.addgene.org/60226/
- AAV:ITR-U6-sgRNA(Kras)-U6-sgRNA(p53)-U6-sgRNA(Lkb1)-pEFS-Rluc-2A-Cre-shortPA-KrasG12D_HDRdonor-ITR (AAV-KPL); Plasmid #60224; http://www.addgene.org/60224/
- BK- pAAV-TRE – SpCas9-NLS-Poly(A)
- BK- pAAV-TRE- SaCas9-NLS-Poly(A)
- BK- pAAV-TRE – dSpCas9-NLS-Poly(A)
- BK- pAAV-TRE – dSaCas9-NLS-Poly(A)
- BK- pAAV-TRE – SpCas9Nickase-NLS-Poly(A)
- BK- pAAV-TRE – SaCas9Nickase-NLS-Poly(A)
CRISPR/CAS9-Based Gene Editing
The IPRD Viral Vector and Gene Editing Cores deliver advanced solutions for biomedical research and therapeutic development. Our expertise in CRISPR/Cas technologies, viral vector engineering, and customized cell and animal model generation empowers investigators to accelerate discovery and translational applications.
We offer a comprehensive portfolio of services that includes the design and production of CRISPR/Cas vectors, generation of genetically engineered cell lines, and customized editing strategies using the latest technologies such as base editing, prime editing, and epigenome editing. Our team is here to partner with you—whether you are developing basic research tools, building disease models, or pursuing therapeutic targets.
- Gene Knockout Cell Line Services
Our core provides fully customizable CRISPR/Cas9-based knockout services in a wide range of mammalian cell lines, including difficult-to-transfect and tumor-derived lines. We deliver functionally validated, long-term stable KO models to researchers at Florida State University and to institutions and companies worldwide.
Our platform includes:
- High-throughput gRNA screening and optimized RNP delivery
- Single- and multi-gene knockout strategies
- Targeted fragment deletions
- End-to-end support from gRNA design to functional validation
Standard workflow:
- Host Cell Characterization
Includes clonability assays, antibiotic resistance profiling, and optimization of transfection/transduction. - gRNA Design & KO Vector Construction
Guides are designed to target conserved exons or critical functional domains of the gene-of-interest. - Transfection & Transduction
Using optimized techniques and house-developed lentiviral and adeno-associated vectors. - Clone Screening
Selection via antibiotic resistance or FACS; expansion of monoclonal or polyclonal populations. - Validation
Characterization via Western blotting, qPCR, ELISA, or reporter assays.
- Specialized Gene Editing Services
Multiplexed KnockoutsGenerate cell lines with multiple gene deletions using pooled or sequential editing strategies.
Disease ModelingCreate precise genetic alterations to model rare or common human diseases in vitro.
Custom CRISPR LibrariesDesign and produce small- to large-scale gRNA libraries for functional genomics studies.
Nuclease Activity ValidationQuantify and optimize CRISPR nuclease performance using a range of molecular and cellular assays.
- Base Editing and Prime Editing
We offer advanced genome editing platforms based on base editors and prime editors—technologies that allow precise, single-nucleotide changes without introducing double-stranded breaks.
Base Editing
Our cytidine and adenine base editors enable targeted C-to-T or A-to-G conversions with minimal off-target effects. These editors are delivered using viral vectors customized for your cell type and application.
- Editing window: 6–10 nucleotides
- Delivery via AAV, lentivirus, or electroporation
- Applications: point mutation correction, functional SNP studies, regulatory region targeting
Prime Editing
We design and deliver pegRNA constructs that enable precise insertions, deletions, or substitutions.
- pegRNA components: spacer, scaffold, reverse transcription template (RTT), primer binding site (PBS)
- nCas9-reverse transcriptase system enables templated DNA repair
- Suitable for applications requiring programmable, high-fidelity edits
With over 15 years of experience in guide RNA design, viral delivery, and construct engineering, we support projects across a wide range of biological systems.
- Epigenome Editing
Epigenetic modifications enable control of gene expression without altering the underlying DNA sequence. We offer full-service epigenome editing platforms using dCas9-based tools fused to:
- DNA methyltransferases and demethylases
- Histone acetyltransferases and deacetylases
Workflow includes:
- Target Identification – Defining loci relevant to gene regulation or disease
- Tool Construction – Building custom dCas9 fusion proteins
- Vector Development – Cloning into expression vectors for delivery
- Delivery – Viral, electroporation, or nanoparticle-based methods
- Selection & Expansion – Isolation of modified clones
- Validation – ChIP, bisulfite sequencing, and gene expression profiling
- Vector and Construct Resources
We maintain an extensive collection of >3,000 in-house optimized CRISPR/Cas constructs for gene knockout, base editing, and more. These are:
- Available directly from the core
- Deposited in the Addgene repository ( https://www.addgene.org )
- Linked to the NGVB repository ( https://ngvbcc.org/ReagentRepository )
Our design and production workflows are supported by 10 patent applications and numerous peer-reviewed publications and funded grants.
Let’s CollaborateWe welcome collaborations with academic, government, and industry partners. Whether your goal is to explore gene function, build disease models, or develop precision therapeutics, our core is ready to support your success.
Contact us today to discuss your project needs and receive a customized consultation.
How to Order
To connect with IPRD’s Viral Vector & Gene Editing Core and place an order, simply email your request to viral.core@med.fsu.edu
