Antisense oligonucleotides selected by hybridisation to scanning arrays are effective reagents in vivo.
Transcripts representing mRNAs of three Xenopus cyclins, B1, B4 and B5, were hybridised to arrays of oligonucleotides scanning the first 120 nt of the coding region to assess the ability of the immobilised oligonucleotides to form heteroduplexes with their targets. Oligonucleotides that produced high heteroduplex yield and others that showed little annealing were assayed for their effect on translation of endogenous cyclin mRNAs in Xenopus egg extracts and their ability to promote cleavage of cyclin mRNAs in oocytes by RNase H. Excellent correlation was found between antisense potency and affinity of oligonucleotides for the cyclin transcripts as measured by the array, despite the complexity of the cellular environment.
Sohail
M
M
Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK. msohail@bioch.ox.ac.uk
Hochegger
H
H
Klotzbücher
A
A
Guellec
R L
RL
Hunt
T
T
Southern
E M
EM
D000818:Animals; D020029:Base Pairing; D001665:Binding Sites; D002457:Cell Extracts; D003001:Cloning, Molecular; D016213:Cyclins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D005786:Gene Expression Regulation; D007106:Immune Sera / Q000276:immunology; D007700:Kinetics; D008958:Models, Molecular; D023282:Nuclease Protection Assays; D009693:Nucleic Acid Hybridization; D020319:Oligodeoxyribonucleotides, Antisense / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D011233:Precipitin Tests; D014176:Protein Biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism; D013379:Substrate Specificity; D014982:Xenopus laevis / Q000235:genetics*
D002457:Cell Extracts; D016213:Cyclins; D007106:Immune Sera; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D016914:Ribonuclease H