Plasmids
Distributed through Addgene, unless otherwise noted.

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.

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

4. TAZ gRNA plasmid (for positive control)
Send request to wpu [at] pulab [dot] org

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).