"ABSTRAK
"Lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV sebagai alternatif agen transfeksi nonviral yang baru telah berhasil disintesis dan diuji karakteristiknya. Penelitian dilakukan dengan tujuan untuk mengetahui karakteristik lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV dalam membentuk kompleks dengan DNA, mengondensasikan DNA, melindungi DNA dari degradasi enzim DNAse, serta mengetahui sitotoksisitas dan kemampuan transfeksinya pada galur sel mamalia, CHO-K1 dan HepG2. Karakteristik lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV kemudian dibandingkan dengan karakteristik dua agen transfeksi komersil, yaitu Poly-L-Lysine dan Lipofectamine 2000. Hasil penelitian menunjukkan bahwa lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV mampu membentuk kompleks dengan DNA pada rasio muatan 0,7, melindungi DNA dari degradasi DNAse pada rasio muatan 1,5, serta memiliki nilai IC50 sebesar 2,940 x 10-2 ?M pada sel CHO-K1 dan 1,964 x 10-2 ?M pada sel HepG2. Namun, lipopeptida tersebut kurang baik dalam mengondensasikan DNA dibandingkan Poly-L-Lysine dan Lipofectamine 2000. Berdasarkan hasil uji transfeksi, lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV telah berhasil mentransfeksikan plasmid pCSII-EF-AcGFP ke dalam sel CHO-K1 dan HepG2. Keberhasilan tersebut ditandai dengan adanya pendaran hasil ekspresi Green Fluorescent Protein pada kedua sel. Meskipun demikian, pendaran yang dihasilkan dinilai kurang maksimal dibandingkan dengan hasil transfeksi oleh Lipofectamine 2000. Hasil transfeksi lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV yang optimal berada pada rasio muatan 7 dengan waktu inkubasi 48 jam.
ABSTRACT
Lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV as a novel nonviral transfection agent has been successfully synthesized and tested by its characteristics. The study aimed to determine the characteristics of lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV in forming complexes with DNA, condensing and protecting DNA from degradation of DNAse enzymes, as well as to reveal its cytotoxicity and ability as a transfection agent in mammalian cell lines, CHO K1 and HepG2. The characteristics of lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV were compared with the characteristics of two commercial transfection agents, Poly L Lysine and Lipofectamine 2000. The result has showed that lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV is able to complex DNA at charge ratio 0.7, protect DNA from DNAse degradation at charge ratio 1.5, and has an IC50 value of 2.940 x 10 2 M in CHO K1 and 1.964 x 10 2 M in HepG2 cells. However, the lipopeptide is not better in condensing the DNA than Poly L Lysine and Lipofectamine 2000. Furthermore, based on the transfection test result, lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV has successfully transfected pCSII EF AcGFP plasmids into CHO K1 and HepG2 cells. The accomplishment is depicted by presence of Green Fluorescent Protein expression in both cells. However, the fluorescence of transfection by lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV is not maximal, compare to Lipofectamine 2000. The optimal transfection result of lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV is at charge ratio 7 and 48 hours incubation time. |
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