Plasmids for Stem Cell Research

Stem cells are proliferative, unspecialized cells with the capacity to differentiate into many different cell types in the body during development. Unlike other cell types, each stem cell has the potential to either remain a stem cell or become another type of cell with a more specialized function. They also have the unique potential to continuously divide while maintaining an undifferentiated state. Stem cells are classified as pluripotent or multipotent, based on their ability to generate different cell lineages. Pluripotent stem cells, such as embryonic cells, can potentially give rise to any other cell type in the body. Multipotent stem cells, such as adult (or somatic) stem cells, can differentiate into multiple cell types, but are typically limited to related cell lineages.

Pluripotent embryonic and multipotent adult stem cells have been studied for over 30 years. All of this research led to Kazutoshi Takahashi and Shinya Yamanaka’s breakthrough development of induced pluripotent stem cells (iPSCs) in both mouse and human cells in 2006 and 2007, respectively. To create iPSCs, fully-differentiated adult somatic cells are returned to a pluripotent state using a cocktail of factors (Oct3/4, Sox2, c-Myc, and Klf4) that are known to maintain pluripotency during embryonic development. This “reprogramming” of the cell to an embryonic state holds immense potential for drug development, disease modeling, and regenerative medicine.

iPSCs, much like their ES cell cousins, have the hallmarks of pluripotent cells--they are capable of differentiating into every other cell type in the body and have the capacity to give rise to an entire organism. iPSC technology removes the possible ethical concerns related to embryonic stem cell use and has the major advantage that iPSCs can potentially be derived from patient-specific or disease-specific cells. For this reason, iPSCs have become an excellent unlimited stem cell source for therapeutic use.

Since 2006, scientists have developed many approaches to generate iPSCs. The generation of iPSCs is relatively simple in concept: ectopically express a cocktail of stem cell reprogramming factors and wait for cells to de-differentiate. However it may be difficult to decide which method to use. Addgene's blog post Delivery Methods for Generating iPSCs provides an overview of several different reprogramming methods with the goal of helping readers choose a strategy suited to their research. Once you know which method you would like to use, browse Addgene's collection of Reprogramming Plasmids in the table below.

Once iPSCs have been created they can be directly differentiated into specific somatic stem cells or fully differentiated cell types. Browse Addgene's collection of Differentiation Plasmids below.

Alternatively, it is also possible to directly differentiate one differentiated somatic cell into another via transdifferentiation. This method of cell lineage conversion can be faster, bypasses intermediate pluripotent stages, and can occur without cell division, all of which lower the risk of mutation and minimize tumorigenicity, an inherent disadvantage of iPS cell technology. Browse Addgene's collection of Transdifferentiation Plasmids below.

Many of the techniques for reprogramming, differentiation, and transdifferentiation utilize plasmids to increase expression of key factors. The tables below will help you find plasmids available from Addgene that can be used to create iPSCs or another cell type. You can also search the iPSC Neurodegenerative Disease Initiative (iNDI) Collection for plasmids that can be used to create cell lines with endogenously-tagged gene variants or browse Addgene’s entire collection of Stem Cell Research Plasmids. For more information on stem cells, visit the NIH Stem Cell information page.

Reprogramming Plasmids

Browse the table below for plasmids used to create induced pluripotent stem cells (iPSCs). Sort the table by delivery method, species, or PI and then select the article link to find more information.

Delivery Method	Species	Description	Article	PI
Lentivirus	Human	Expression of human Oct4, Sox2, Klf4, Myc, and Brd3R from five separate lentiviral plasmids	The acetyllysine reader BRD3R promotes human nuclear reprogramming and regulates mitosis. Nat Commun. 2016 Mar 7;7:10869.	Hu
CRISPRa	Human	CRISPR activator system for reprogramming human cells to pluripotency by activating enogenous genes	Human pluripotent reprogramming with CRISPR activators. Nat Commun. 2018 Jul 6;9(1):2643.	Otonkoski
Lentivirus	Human	Expression of human Sox2, Nanog, Oct4, and Lin28 from four separate lentiviral plasmids	Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells. Science. 2007 Dec 21;318(5858):1917-20.	Thomson
Lentivirus	Human	Doxycycline-inducible expression of human Oct4, Sox2, Klf4, and c-Myc from four separate lentiviral plasmids	A drug-inducible system for direct reprogramming of human somatic cells to pluripotency. Cell Stem Cell. 2008 Sep 11. 3(3):346-53.	Jaenisch
Lentivirus	Human	Single polycistronic lentiviral vector for the expression of human Oct4, Sox2, and Klf4	Polycistronic lentiviral vector for "hit and run" reprogramming of adult skin fibroblasts to induced pluripotent stem cells. Stem Cells. 2009 May . 27(5):1042-9.	Townes
Lentivirus	Human	Single polycistronic lentiviral vector for the expression of human Oct4, Klf4, Sox2, and c-Myc	Human Induced Pluripotent Stem Cells Produced Under Xeno-Free Conditions. Stem Cells Dev. 2010 Aug;19(8):1221-9.	Cibelli
Lentivirus	Human	Single polycistronic, doxycycline-inducible lentiviral vector for the expression of human Oct4, Klf4, c-Myc, and Sox2	Integrative Analyses of Human Reprogramming Reveal Dynamic Nature of Induced Pluripotency. Cell. 2015 Jul 16;162(2):412-24.	Mikkelsen
Lentivirus	Human	Expression of human Klf4, Oct4, c-Myc, and Sox2 as VP-16 transcriptional activating fusions in single lentiviral vectors for human iPSC generation	OCT4 and SOX2 Work as Transcriptional Activators in Reprogramming Human Fibroblasts. Cell Rep. 2017 Aug 15;20(7):1585-1596.	Ptashne
Minicircle	Human	Non-integrating mini-intronic plasmid (MIP) polycistronic expression of human Oct4, Klf4, Sox2, c-Myc and hairpin RNA p53 for single plasmid reprogramming	Novel codon-optimized mini-intronic plasmid for efficient, inexpensive, and xeno-free induction of pluripotency. Sci Rep. 2015 Jan 28;5:8081.	Kay and Wu
MMLV-derived Retrovirus	Human	Yamanaka factors for generating human iPS cells; retroviral expression of human Sox2, Oct3/4, Klf4, and c-Myc from four separate plasmids	Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007 Nov 30. 131(5):861-72.	Yamanaka
Plasmid	Human	Non-integrating transient expression of human L-Myc, Lin28, Sox2, Klf4, Oct3/4, Glis1, and EBNA1 from separate plasmids	An Efficient Non-viral Method to Generate Integration-Free Human iPS Cells from Cord Blood and Peripheral Blood Cells. Stem Cells. 2012 Nov 29.	Yamanaka
Replicating EBNA1 episome	Human	Non-integrating EBNA1-mediated polycistronic expression of human Sox2, KLF4, L-Myc, Lin28, OCT3/4, and shRNA against p53 in different gene/insert combinations	A more efficient method to generate integration-free human iPS cells. Nat Methods. 2011 May;8(5):409-12.	Yamanaka
Replicating EBNA1 episome	Human	Non-integrating polycistronic expression of human Oct4, Klf4, Sox2, c-Myc, Nanog, Lin28, NR5A2, and microRNA 302/367 in three different combinations of reprogramming factors	Efficient germ-line transmission obtained with transgene-free induced pluripotent stem cells. Proc Natl Acad Sci U S A. 2014 Jul 22;111(29):10678-83.	Capecchi
Replicating EBNA1 episome	Human	Fluorescent-tagged EBNA1-mediated expression of human Oct3/4, shp53, Klf4, Sox2, L-Myc, Lin28 from separate non-integrating episomes to allow sorting and dosage tracking of reprogramming factors	Fluorescent tagged episomals for stoichiometric induced pluripotent stem cell reprogramming. Stem Cell Res Ther. 2017 Jun 5;8(1):132.	Zovein
Replicating EBNA1 episome	Human	Non-integrating EBNA1-mediated polycistronic expression of human Oct4, Sox2, Myc, Klf4, BCL2L1, and EBNA1 from three separate plasmids	A Facile Method to Establish Human Induced Pluripotent Stem Cells From Adult Blood Cells Under Feeder-Free and Xeno-Free Culture Conditions: A Clinically Compliant Approach. Stem Cells Transl Med. 2015 Mar 5. pii: sctm.2014-0214.	Cheng
RNA	Human	Reprogramming factors for delivering synthetic mRNAs encoding human Klf4, Oct4, Sox2, c-Myc, and Lin28A to somatic cells from separate plasmids	Highly Efficient Reprogramming to Pluripotency and Directed Differentiation of Human Cells with Synthetic Modified mRNA. Cell Stem Cell. 2010 Nov 5;7(5):618-30.	Rossi
RNA	Human	Non-integrating, polycistronic, self-replicating VEE RNA species expressing human Oct4, Klf4, Sox2, with either c-Myc or Glis1	Efficient generation of human iPSCs by a synthetic self-replicative RNA. Cell Stem Cell. 2013 Aug 1;13(2):246-54.	Dowdy
Adenovirus	Mouse	Non-integrating expression of mouse Sox2, Oct4, c-Myc, and Klf4 from four separate adenoviral plasmids	Induced Pluripotent Stem Cells Generated Without Viral Integration. Science. 2008 Nov 7;322(5903):945-9.	Hochedlinger
Lentivirus	Mouse	Doxycycline-inducible expression of mouse Oct4, Sox2, Klf4, and c-Myc from four separate lentiviral plasmids	Sequential expression of pluripotency markers during direct reprogramming of mouse somatic cells. Cell Stem Cell. 2008 Feb 7. 2(2):151-9.	Jaenisch
Lentivirus	Mouse	Polycistronic, doxycycline-inducible lentiviral vectors for the expression of mouse Oct4, Klf4, c-Myc, and Sox2	Reprogramming of murine and human somatic cells using a single polycistronic vector. Proc Natl Acad Sci U S A. 2009 Jan 6. 106(1):157-62.	Jaenisch
Lentivirus	Mouse	Doxycycline-inducible lentiviral expression of mouse Oct4, Sox2, Klf4, and c-Myc from four separate plasmids	Defining molecular cornerstones during fibroblast to iPS cell reprogramming in mouse. Cell Stem Cell. 2008 Mar 6. 2(3):230-40.	Hochedlinger
MMLV-derived Retrovirus	Mouse	Original set of Yamanaka factors for generating mouse iPS cells; retroviral expression of mouse Sox2, Oct3/4, Klf4, and c-Myc from separate plasmids	Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006 Aug 25. 126(4):663-76.	Yamanaka
piggyBac	Mouse	Doxycycline-inducible piggyBac transposon reprogramming system; expression of mouse Oct4, Sox2, Klf4, and c-Myc from polycistronic cassettes	KLF4 N-terminal variance modulates induced reprogramming to pluripotency. Stem Cell Reports. 2015 Apr 14;4(4):727-43.	Woltjen
Plasmid	Mouse	Non-integrating polycistronic expression of mouse Oct4, Klf4, Sox2, and c-Myc from two separate plasmids	Generation of Mouse Induced Pluripotent Stem Cells Without Viral Vectors. Science. 2008 Nov 7;322(5903):949-53.	Yamanaka
Replicating EBNA1 episome	Mouse	Non-integrating EBNA1-mediated polycistronic expression of mouse Oct4, Sox2, Klf4, c-Myc, and Lin28	Efficient human iPS cell derivation by a non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures. Cell Res. 2011 Mar;21(3):518-29.	Cheng
piggyBac	Mouse	Doxycycline-inducible piggyBac transposon reprogramming system; expression of mouse Oct4, Sox2, Klf4, and c-Myc from polycistronic cassettes.	piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells. Nature. 2009 Apr 9;458(7239):766-70.	Nagy
Lentivirus	Mouse	Polycistronic, doxycycline-inducible lentiviral vectors for the expression of mouse Oct4, Klf4, c-Myc, and Sox2	Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental Potential of iPSCs. Cell Stem Cell. 2019 Oct 30. pii: S1934-5909(19)30423-0.	Schöler

You can also find gene-specific plasmids at the following links: NANOG, OCT4, SOX2, MYC, KLF4, LIN28.

Differentiation and Transdifferentiation Plasmids

Browse the table below for plasmids used for differentiation of iPSCs or transdifferentiation of a somatic cell to a new type of somatic cell. Sort the table by cell type, delivery system, species, or PI and then select the article link to find more information.

Convert From	Convert To	Delivery Method	Species	Article	PI
iPSCs	Cardiomyocytes	Lentiviral	Human	Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci U S A. 2012 May 29.	Palecek
Fibroblasts	Cardiomyocytes	Lentiviral	Mouse	Optimization of direct fibroblast reprogramming to cardiomyocytes using calcium activity as a functional measure of success. Mol Cell Cardiol. 2013 Apr 13;60C:97-106.	Gearhart
Fibroblasts	Hematopoietic Progenitor Cells	Lentiviral	Mouse	Direct reprogramming of murine fibroblasts to hematopoietic progenitor cells. Cell Rep. 2014 Dec 11;9(5):1871-84.	Lacaud
Fibroblasts	Hematopoietic Progenitor Cells	Lentiviral	Human	Cooperative Transcription Factor Induction Mediates Hemogenic Reprogramming. Cell Rep. 2018 Dec 4;25(10):2821-2835.e7.	Pereira
Fibroblasts	Sertoli Cells	Lentiviral	Mouse	Direct Reprogramming of Fibroblasts into Embryonic Sertoli-like Cells by Defined Factors. Cell Stem Cell. 2012 Sep 7;11(3):373-86.	Jaenisch
Fibroblasts	Hepatocytes	Retroviral	Mouse	Direct conversion of mouse fibroblasts to hepatocyte-like cells by defined factors. Nature. 2011 Jun 29;475(7356):390-3.	Suzuki
Pancreatic Exocrine Cells	Beta-cells	Adenoviral	Mouse	In vivo reprogramming of adult pancreatic exocrine cells to beta-cells. Nature. 2008 Aug 27.	Melton
Pancreatic Acinar Cells	induced Beta Cells	Adenoviral	Mouse	Long-term persistence and development of induced pancreatic beta cells generated by lineage conversion of acinar cells. Nature Biotechnology. 2014 Dec;32(12):1223-30.	Zhou
Astrocytes	Dopaminergic Neurons	Lentiviral	Human	Efficient conversion of astrocytes to functional midbrain dopaminergic neurons using a single polycistronic vector. PLoS One. 2011;6(12):e28719.	Gearhart
Fibroblasts	Neurons	Lentiviral	Mouse	Direct conversion of fibroblasts to functional neurons by defined factors. Nature. 2010 Feb 25. 463(7284):1035-41.	Wernig
Fibroblasts	Dopaminergic Neurons	Lentiviral	Human	Direct conversion of human fibroblasts to dopaminergic neurons. Proc Natl Acad Sci U S A. 2011 Jun 21;108(25):10343-8.	Parmar
Fibroblasts	Neurons	Lentiviral	Human	MicroRNA-mediated conversion of human fibroblasts to neurons. Nature. 2011 Jul 13. doi: 10.1038/nature10323.	Crabtree
Fibroblasts	Motor Neurons	Retroviral	Mouse/Human	Conversion of mouse and human fibroblasts into functional spinal motor neurons. Cell Stem Cell. 2011 Sep 2;9(3):205-18.	Eggan
Fibroblasts	Neural Precursor Cells	Lentiviral	Mouse	Direct conversion of mouse fibroblasts to self-renewing, tripotent neural precursor cells. Proc Natl Acad Sci U S A. 2012 Feb 14;109(7):2527-32.	Wernig
Fibroblasts	Neural Stem Cells	Retroviral	Mouse	Direct reprogramming of fibroblasts into neural stem cells by defined factors. Cell Stem Cell. 2012 Apr 6;10(4):465-72.	Schöler
Adult Skin Fibroblasts	Cholinergic Neurons	Lentiviral	Human	Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic neurons. Nat Commun. 2013;4:2183.	Zhang
Fetal Lung Fibroblasts	Cholinergic Neurons	Lentiviral	Human	Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic neurons. Nat Commun. 2013;4:2183.	Zhang
Fibroblasts	Oligodendrocyte Precursor Cells	Lentiviral	Mouse/Rat	Generation of oligodendroglial cells by direct lineage conversion. Nat Biotechnol. 2013 May;31(5):434-9.	Wernig
Fibroblasts	Oligodendrocyte Progenitor Cells	Lentiviral	Mouse	Transcription factor-mediated reprogramming of fibroblasts to expandable, myelinogenic oligodendrocyte progenitor cells. Nat Biotechnol. 2013 May;31(5):426-33.	Tesar
Fibroblasts	Medium Spiny Neurons	Lentiviral	Human	Generation of Human Striatal Neurons by MicroRNA-Dependent Direct Conversion of Fibroblasts. Neuron. 2014 Oct 22;84(2):311-23.	Yoo
Fibroblasts	Neurons	Lentiviral	Human	Direct neural conversion from human fibroblasts using self-regulating and nonintegrating viral vectors. Stem Cell Reports. 2014 Nov 6. pii: S2213-6711(14)00307-5.	Jaenisch
Fibroblasts	Neural Stem Cells	Lentiviral	Mouse	Direct Lineage Conversion of Adult Mouse Liver Cells and B Lymphocytes to Neural Stem Cells. Cell Reports. 2014 Dec 11;9(5):1673-80.	Parmar
Fibroblasts	Astrocytes	Lentiviral	Mouse	Direct conversion of fibroblasts into functional astrocytes by defined transcription factors. Stem Cell Reports. 2015 Jan 13;4(1):25-36.	Broccoli
Fibroblasts	Sensory Neurons	Lentiviral	Mouse	Selective conversion of fibroblasts into peripheral sensory neurons. Nat Neurosci. 2015 Jan;18(1):25-35.	Baldwin
Fibroblasts	iTSCs	Lentiviral	Mouse	Extensive Nuclear Reprogramming Underlies Lineage Conversion into Functional Trophoblast Stem-like Cells. Cell Stem Cell. 2015 Nov 5;17(5):543-56.	Buganim
Skin Fibroblasts	Motor Neurons	Lentiviral	Human	Direct Lineage Reprogramming Reveals Disease-Specific Phenotypes of Motor Neurons from Human ALS Patients. Cell Rep. 2016 Jan 5;14(1):115-28.	Zhang
Adult Dermal Fibroblasts	Neurons	Lentiviral	Human	REST suppression mediates neural conversion of adult human fibroblasts via microRNA-dependent and -independent pathways. EMBO Mol Med. 2017 Aug;9(8):1117-1131.	Parmar
Fibroblasts	Dentritic Cells	Lentiviral	Mouse	Direct reprogramming of fibroblasts into antigen-presenting dendritic cells. Sci Immunol. 2018 Dec 7;3(30). pii: 3/30/eaau4292.	Pereira
Fibroblasts	Neurons	Lentiviral	Human	Chemical modulation of transcriptionally enriched signaling pathways to optimize the conversion of fibroblasts into neurons. Elife. 2019 May 17;8. pii: 41356.	Gage
Fibroblasts	iPSCs, iTSCs, and iXENs	Lentiviral	Mouse	Direct Induction of the Three Pre-implantation Blastocyst Cell Types from Fibroblasts. Cell Stem Cell. 2019 Jun 6.	Buganim
Glial Cells	Retinal Ganglion Cells	AAV/CRISPR	Mouse	Glia-to-Neuron Conversion by CRISPR-CasRx Alleviates Symptoms of Neurological Disease in Mice. Cell. 2020 Apr 30;181(3):590-603.e16.	Yang
iPSCs	Bipolar Neurons	Lentiviral	Human	Rapid neurogenesis through transcriptional activation in human stem cells. Mol Syst Biol. 2014 Nov 17;10(11):760.	Weiss
iPSCs & iPSC-NPCs	Neurons	Lentiviral	Human	Rapid Ngn2-induction of excitatory neurons from hiPSC-derived neural progenitor cells. Methods. 2015 Nov 25. pii: S1046-2023(15)30159-6.	Brennand
iPSCs	Cortical or Lower Motor Neurons	CRISPR/TALEN	Human	Transcription Factor-Mediated Differentiation of Human iPSCs into Neurons. Curr Protoc Cell Biol. 2018 Jun;79(1):e51. Next generation vectors for transcription factor mediated differentiation of iPSCs into cortical and motor neurons. Unpublished.	Ward
Glial Progenitor Cells	GABAergic Interneurons	Lentiviral	Human	Direct Conversion of Human Stem Cell-Derived Glial Progenitor Cells into GABAergic Interneurons. Cells. 2020 Nov 10;9(11). pii: cells9112451. doi: 10.3390/cells9112451.	Ottosson

Stem Cell Research Plasmids

This table contains a general list of plasmids that have been used in Stem Cell research. You can search or sort this table based on plasmid name, species, gene name, and more.