TY - JOUR
T1 - High-efficiency transfection of nucleic acids by the modified calcium phosphate precipitation method in chondrocytes
AU - Qureshi, Hamid Yaqoob
AU - Ahmad, Rasheed
AU - Zafarullah, Muhammad
PY - 2008/11/15
Y1 - 2008/11/15
N2 - Extracellular matrix (ECM)-rich cartilage-derived chondrocytes are difficult to transfect with DNA/RNA. We modified the classical calcium phosphate transfection method by detaching adherent chondrocytes with trypsin and resuspending in CaPo4-nucleic acid precipitate followed by readherence for 24 h. Due to the absence of ECM, chondrocytes could be transfected with 80% efficiency. Potent gene silencing with several antisense oligonucleotides and small interfering RNAs and strong promoter-luciferase activity could be achieved. This approach is applicable to any adherent or suspended cells and has utility in gene knockdown, ectopic overexpression, promoter regulation studies, and gene delivery in tissue engineering and gene therapy applications.
AB - Extracellular matrix (ECM)-rich cartilage-derived chondrocytes are difficult to transfect with DNA/RNA. We modified the classical calcium phosphate transfection method by detaching adherent chondrocytes with trypsin and resuspending in CaPo4-nucleic acid precipitate followed by readherence for 24 h. Due to the absence of ECM, chondrocytes could be transfected with 80% efficiency. Potent gene silencing with several antisense oligonucleotides and small interfering RNAs and strong promoter-luciferase activity could be achieved. This approach is applicable to any adherent or suspended cells and has utility in gene knockdown, ectopic overexpression, promoter regulation studies, and gene delivery in tissue engineering and gene therapy applications.
UR - http://www.scopus.com/inward/record.url?scp=52249098833&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2008.07.027
DO - 10.1016/j.ab.2008.07.027
M3 - Article
C2 - 18703012
AN - SCOPUS:52249098833
VL - 382
SP - 138
EP - 140
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 2
ER -