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Plasmid-Reference_book.png 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.

Schematic of reprogramming stem cells

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. 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
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

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 transdifferntion 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.

From → To Delivery Method Species Article PI
iPSC → Cardiomyocyte 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
iPSC → Bipolar Neurons Lentiviral Human Rapid neurogenesis through transcriptional activation in human stem cells. Mol Syst Biol. 2014 Nov 17;10(11):760. Weiss
iPSC → 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. Ward
Fibroblast → 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
Fibroblast → Neural Stem Cell Retroviral Mouse Direct reprogramming of fibroblasts into neural stem cells by defined factors. Cell Stem Cell. 2012 Apr 6;10(4):465-72. Scholer
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
Fibroblast → Neurons Lentiviral Mouse Direct conversion of fibroblasts to functional neurons by defined factors. Nature. 2010 Feb 25. 463(7284):1035-41 Wernig
Fibroblast → Neural Stem Cell Lentiviral Mouse Direct Lineage Conversion of Adult Mouse Liver Cells and B Lymphocytes to Neural Stem Cells. Stem Cell Reports. 2014 Nov 6. pii: S2213-6711(14)00307-5. Jaenisch
Fibroblast → Sensory Neurons Lentiviral Mouse Selective conversion of fibroblasts into peripheral sensory neurons. Nat Neurosci. 2015 Jan;18(1):25-35. Baldwin
Fibroblast → Astrocyte Lentiviral Mouse Direct conversion of fibroblasts into functional astrocytes by defined transcription factors. Stem Cell Reports. 2015 Jan 13;4(1):25-36. Broccoli
Fibroblast → Oligodendrocyte Progenitor Lentiviral Mouse Transcription factor-mediated reprogramming of fibroblasts to expandable, myelinogenic oligodendrocyte progenitor cells. Nat Biotechnol. 2013 May;31(5):426-33. Tesar
Fibroblast → Cardiomyocyte 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
Fibroblast → Sertoli Cell 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
Fibroblast → Hematopoietic Progenitor Lentiviral Mouse Direct reprogramming of murine fibroblasts to hematopoietic progenitor cells. Cell Rep. 2014 Dec 11;9(5):1871-84. Lacaud
Pancreatic Exocrine → Beta-cell Adenoviral Mouse In vivo reprogramming of adult pancreatic exocrine cells to beta-cells. Nature. 2008 Aug 27. Melton
Skin Fibroblast → Motor Neuron 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 Skin Fibroblast → Cholinergic Neuron Lentiviral Human Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic neurons. Nat Commun. 2013;4:2183. Zhang
Fetal Lung Fibroblast → Cholinergic Neuron Lentiviral Human Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic neurons. Nat Commun. 2013;4:2183. Zhang
Fibroblast → Dopaminergic Neuron 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
Adult Dermal Fibroblast → Neuron 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
Astrocytes → Dopaminergic Neuron Lentiviral Human Efficient conversion of astrocytes to functional midbrain dopaminergic neurons using a single polycistronic vector. PLoS One. 2011;6(12):e28719. Gearhart
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