Plasmids
Distributed through Addgene, unless otherwise noted.
Link to Pu lab plasmids at Addgene.

CASAAV cardiomyocyte somatic gene knockout
  • Materials for: Guo, Y., VanDusen, N. J., Zhang, L., Gu, W., Sethi, I., Guatimosim, S., Ma, Q., Jardin, B. D., Ai, Y., Zhang, D., Chen, B., Guo, A., Yuan, G. C., Song, L. S. & Pu, W. T. Analysis of Cardiac Myocyte Maturation Using CASAAV, A Platform for Rapid Dissection of Cardiac Myocyte Gene Function In Vivo. Circ Res (2017).

1. AAV-U6gRNA1-U6gRNA2-TnT-Cre plasmid.
Expresses two guide RNAs from U6 promoters and Cre from the cardiac Tnnt2 promoter.
Available from addgene (#87682).

Genome Editing

  • Materials for: Wang, G., Yang, L., Grishin, D., Rios, X., Ye, L. Y., Hu, Y., Li, K., Zhang, D., Church, G. M. & Pu, W. T. Efficient, footprint-free human iPSC genome editing by consolidation of Cas9/CRISPR and piggyBac technologies. Nat Protoc 12, 88-103 (2017):

1. pPB-rtTA-hCas9-puro-PB plasmid. map
The plasmid was made by the Church lab. Please contact hms_materialtransfer [at] harvard [dot] edu to complete an MTA. Also include in the email the shipping address. The material will be provided by way of the Pu lab. Recipient will be responsible for shipping costs.

2. Excision only piggyBac transposase expression vector
System Biosciences, cat. no. PB220PA-1

3. Super piggyBac transposase expression vector
System Biosciences, cat. no. PB210PA-1


pEntr-flbio
Vector for creating flbio-tagged cDNAs in the gateway system
  • He, A. and Pu, W. T. (2010) Curr Protoc Mol Biol Chapter 21, Unit 21.20
  • He, A., Kong, S.W., Ma, Q., and Pu, W.T. 2011. Co-occupancy by multiple cardiac transcription factors identifies transcriptional enhancers active in heart. Proc Natl Acad Sci U S A. 108:5632-5637.

pEntr-GATA4-flbio
pEntr-Mef2a-flbio
pEntr-Tbx5-flbio
pEntr-Srf-flbio
pEntr-Nkx2-5-flbio

Entry plasmids for expression of flbio-tagged cardiac transcription factors.
  • He, A. and Pu, W. T. (2010) Curr Protoc Mol Biol Chapter 21, Unit 21.20
  • He, A., Kong, S.W., Ma, Q., and Pu, W.T. 2011. Co-occupancy by multiple cardiac transcription factors identifies transcriptional enhancers active in heart. Proc Natl Acad Sci U S A. 108:5632-5637.

pEntr-3xFLAG-YAP1-S127A
Entry plasmid for expression of 3xFLAG-tagged YAP1 with S127A mutation
  • von Gise, A., Lin, Z., Schlegelmilch, K., Honor, L. B., Pan, G. M., Buck, J. N., Ma, Q., Ishiwata, T., Zhou, B., Camargo, F. D. & Pu, W. T. YAP1, the nuclear target of Hippo signaling, stimulates heart growth through cardiomyocyte proliferation but not hypertrophy. Proc Natl Acad Sci U S A 109, 2394-2399 (2012).

YTIP-GFP
Express peptide that interferes with YAP-TEAD interaction
  • von Gise, A., Lin, Z., Schlegelmilch, K., Honor, L. B., Pan, G. M., Buck, J. N., Ma, Q., Ishiwata, T., Zhou, B., Camargo, F. D. & Pu, W. T. YAP1, the nuclear target of Hippo signaling, stimulates heart growth through cardiomyocyte proliferation but not hypertrophy. Proc Natl Acad Sci U S A 109, 2394-2399 (2012).