A Recombinant Collagen-mRNA Platform for Controllable Protein Synthesis.
We have developed a collagen-mRNA platform for controllable protein production that is intended to be less prone to the problems associated with commonly used mRNA therapy as well as with collagen skin-healing procedures. A collagen mimic was constructed according to a recombinant method and was used as scaffold for translating mRNA chains into proteins. Cysteines were genetically inserted into the collagen chain at positions allowing efficient ribosome translation activity while minimizing mRNA misfolding and degradation. Enhanced green fluorescence protein (eGFP) mRNA bound to collagen was successfully translated by cell-free Escherichia coli ribosomes. This system enabled an accurate control of specific protein synthesis by monitoring expression time and level. Luciferase-mRNA was also translated on collagen scaffold by eukaryotic cell extracts. Thus we have demonstrated the feasibility of controllable protein synthesis on collagen scaffolds by ribosomal machinery.
Department of Biomaterials, College of Materials, Xiamen University, 422, Siming South Road, Xiamen 361005 (China).
Liping
L
Sun
Department of Biomaterials, College of Materials, Xiamen University, 422, Siming South Road, Xiamen 361005 (China).
Yunjing
Y
Xiong
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel).
Anat
A
Bashan
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel).
Ella
E
Zimmerman
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel).
Shirley
S
Shulman Daube
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel).
Yoav
Y
Peleg
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel).
Shira
S
Albeck
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel).
Tamar
T
Unger
Sheba Medical Center, 1 Sheba Street, Tel Hashomer 52621 (Israel).|Sackler School of Medicine, Tel Aviv University, 10 Levanon Street, Tel Aviv 69978 (Israel).
Hagith
H
Yonath
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel).
Miri
M
Krupkin
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel).
Donna
D
Matzov
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel). ada.yonath@weizmann.ac.il.
Ada
A
Yonath
D002474:Cell-Free System / Q000378:metabolism; D003094:Collagen / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D049408:Luminescent Agents / Q000032:analysis / Q000378:metabolism; D058619:Maltose-Binding Proteins / Q000737:chemistry / Q000235:genetics; D014176:Protein Biosynthesis; D055503:Protein Multimerization; D055550:Protein Stability; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics
D049408:Luminescent Agents; D058619:Maltose-Binding Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D003094:Collagen; D008156:Luciferases
collagen; mRNA; protein synthesis; ribosomes