ϸСRNA²âÐò£¨miRNA-Seq£©ÊÇÒ»ÖÖÓÃÓÚÑо¿Ï¸Ð¡RNA£¨miRNA£©±í´ïµÄ¸ßͨÁ¿²âÐòÊÖÒÕ¡£miRNAÊÇÒ»Àà¶ÌСµÄ·Ç±àÂëRNA·Ö×Ó£¬ÔÚ»ùÒòµ÷¿Ø¡¢Ï¸°ûÉúÃüÖÜÆÚ¡¢·¢ÓýºÍ¼²²¡µÈ¶à¸öÉúÎïѧÀú³ÌÖÐÊ©Õ¹Ö÷Òª×÷Óá£
1£©¸ßÖÊÁ¿µÄÊý¾ÝÌìÉú
2£©ÍêÕûµÄÁýÕÖ¹æÄ£
3£©¶àά¶ÈµÄÊý¾ÝÆÊÎö
4£©×¨ÒµµÄÊý¾Ý½â¶Á
5£©±¨¸æÒÔ¼°¶¨ÖÆ»¯µÄЧÀÍ
1£©Êý¾ÝÔ¤´¦Öóͷ££º¶ÔÔʼ²âÐòÊý¾Ý¾ÙÐÐÖÊÁ¿¿ØÖƺ͹ýÂË£¬È¥³ýµÍÖÊÁ¿µÄ¶Á¶Î¡¢ÌÖÂÛÐòÁкÍÖظ´ÐòÁеȡ£Í¬Ê±£¬»¹¿ÉÒÔ¶ÔÊý¾Ý¾ÙÐÐÈ¥³ýrRNAÐòÁеIJÙ×÷£¬ÒÔïÔÌ×ÌÈÅ¡£
2£©ÐòÁбȶԣº½«Ô¤´¦Öóͷ£ºóµÄ²âÐòÊý¾ÝÓëmiRNAÊý¾Ý¿â£¨ÈçmiRBase£©¾ÙÐбȶԣ¬Ê¹ÓñȶԹ¤¾ß£¨ÈçBowtie¡¢BWA¡¢STARµÈ£©¾ÙÐÐÐòÁбȶԡ£ÕâÒ»°ì·¨½«È·¶¨Ã¿¸ö¶Á¶ÎµÄȪԴλÖ㬲¢½«ÆäÓ³Éäµ½miRNAÉÏ¡£
3£©±í´ïÁ¿ÅÌË㣺ƾ֤±È¶ÔЧ¹û£¬Í³¼Æÿ¸ömiRNAµÄ¶Á¶ÎÊý»òÁýÕֶȣ¬À´Ô¤¼ÆÆä±í´ïÁ¿¡£³£ÓõÄÒªÁì°üÀ¨RPKM£¨Reads Per Kilobase of transcript per Million mapped reads£©»òFPKM£¨Fragments Per Kilobase of transcript per Million mapped fragments£©¡£
4£©²î±ð±í´ïÆÊÎö£ºÍ¨¹ý½ÏÁ¿²î±ðÑùÆ·Ö®¼äµÄmiRNA±í´ïÁ¿£¬¿ÉÒÔÅжϲî±ð±í´ïµÄmiRNA¡£³£ÓõIJî±ð±í´ïÆÊÎöÒªÁì°üÀ¨DESeq¡¢edgeRµÈ¡£ÕâЩҪÁì¿ÉÒÔƾ֤ͳ¼Æѧģ×Ó£¬Ê¶±ð²î±ð±í´ïmiRNA²¢ÅÌËãÆäÏÔÖøÐÔ¡£
5£©¹¦Ð§×¢Êͺ͸»¼¯ÆÊÎö£º¶Ô²î±ð±í´ïµÄmiRNA¾ÙÐй¦Ð§×¢ÊÍ£¬¼´½«ÆäÓ³Éäµ½ÒÑÖªmiRNAµÄ¹¦Ð§×¢ÊÍÊý¾Ý¿âÖУ¬ÈçmiRBase¡¢TargetScanµÈ¡£±ðµÄ£¬»¹¿ÉÒÔ¾ÙÐи»¼¯ÆÊÎö£¬Ñ°ÕÒ²î±ð±í´ïmiRNAÔÚÌض¨ÉúÎïÀú³Ì¡¢Í¨Â·»ò¹¦Ð§Éϵĸ»¼¯ÇéÐΡ£
6£©Ð§¹ûÑéÖ¤£ºÍ¨¹ýʵÑéÑéÖ¤²î±ð±í´ïmiRNAµÄÉúÎïѧ¹¦Ð§ºÍµ÷¿Ø»úÖÆ£¬ÀýÈ磬ͨ¹ý¶¨Á¿PCR¡¢Northern blotµÈÒªÁìÑéÖ¤²âÐòЧ¹ûµÄ׼ȷÐԺͿɿ¿ÐÔ¡£
1£©miRNA±í´ïµ÷¿ØÑо¿£º miRNA-Seq¿ÉÒÔÓÃÓÚÆÊÎö²î±ðϸ°ûÀàÐÍ¡¢·¢Óý½×¶Î¡¢ÐÄÀí״̬»ò¼²²¡×´Ì¬ÏÂmiRNAµÄ±í´ïת±ä£¬×ÊÖúÏàʶmiRNAÔÚ»ùÒòµ÷¿ØÖеÄ×÷Óá£
2£©miRNAÓë¼²²¡¹ØÁªÑо¿£º ͨ¹ý½ÏÁ¿¼²²¡»¼ÕߺÍÕý³£±ÈÕÕ×éµÄmiRNA±í´ïÊý¾Ý£¬¿ÉÒÔÅжÏÓë¼²²¡Ïà¹ØµÄmiRNA£¬´Ó¶øÉîÈëÏàʶmiRNAÔÚ¼²²¡±¬·¢ÆøÖÆÖеÄ×÷Óá£
3£©miRNAÓëϸ°ûÖÜÆÚ¡¢µòÍöºÍ·Ö½âÑо¿£º miRNA-Seq¿ÉÒÔ×ÊÖúÏàʶmiRNAÔÚϸ°ûÖÜÆÚ¡¢µòÍöºÍ·Ö½âµÈÉúÎïѧÀú³ÌÖеĹ¦Ð§¡£
4£©miRNAÓë°©Ö¢Ñо¿£º miRNAÔÚ¶àÖÖ°©Ö¢Öбí´ïÒì³££¬miRNA-Seq¿ÉÒÔÕ¹ÏÖ°©Ö¢ÖÐmiRNAµÄ±í´ïת±ä£¬´Ó¶øÏàʶmiRNAÔÚ°©Ö¢Éú³¤ÖеÄ×÷Óá£
5£©miRNAÓëÒ©ÎïÖÎÁÆÑо¿£º ÏàʶmiRNAÔÚ¼²²¡ÖеÄ×÷ÓÃÓÐÖúÓÚÑз¢Õë¶ÔmiRNAµÄÖÎÁÆÕ½ÂÔ£¬ÈçʹÓÃmiRNA×÷ΪÖÎÁưеã»òµ÷¿Ø¼Á¡£
6£©miRNAµ÷¿ØÍøÂçÆÊÎö£º ͨ¹ýmiRNA-SeqÊý¾Ý£¬¿ÉÒÔ¹¹½¨miRNAÓë°Ð»ùÒòÖ®¼äµÄµ÷¿ØÍøÂ磬ÉîÈëÏàʶmiRNAµÄµ÷¿Ø»úÖƺÍ×÷Óá£
7£©miRNAÓëÃâÒßϵͳÑо¿£º miRNAÔÚµ÷ÀíÃâÒßÓ¦´ð¡¢Ñ×Ö¢ºÍÃâÒßϸ°û·Ö½âÖÐÊ©Õ¹Ö÷Òª×÷Óã¬miRNA-SeqÓÐÖúÓÚÑо¿miRNAÔÚÃâÒßϵͳÖеĹ¦Ð§¡£
8£©miRNA×÷ΪÉúÎï±ê¼ÇÎïµÄÓ¦Ó㺠һЩmiRNAÔÚÌض¨¼²²¡×´Ì¬Ï±í´ïÏÔ×Åת±ä£¬¿ÉÒÔ×÷ΪDZÔÚµÄÉúÎï±ê¼ÇÎÓÃÓÚ¼²²¡µÄÕï¶ÏºÍ¼à²â¡£
A fecal miRNA signature by small RNA sequencing accurately distinguishes colorectal cancers: results from a multicentric study£¨Gastroenterology£¬2023£¬IF=33.883£©
ͨ¹ýСRNA²âÐòµÄ·à±ãmiRNA±ê¼Ç׼ȷÇø·Ö½áÖ±³¦°©:À´×Ô¶àÖÐÐÄÑо¿µÄЧ¹û
ÏÖÔÚÓÃÓÚ½áÖ±³¦°©(CRC)ɸ²éµÄ·à±ã¼ì²âÔÚ¼ì²âÔçÆÚÖ×Áö»ò°©Ç°²¡±ä·½ÃæµÄ׼ȷÐÔÓÐÏÞ¡£ÔÚÕâ·½Ã棬×÷ÕßÖÜÈ«ÆÀ¹ÀÁË·à±ãmicroRNA (miRNA)Æ××÷Ϊ·ÇÇÖÈëÐÔ½áÖ±³¦°©Õï¶ÏµÄÉúÎï±ê¼ÇÎï¡£ÔÚ¶à¸öÉúÎï±ê±¾Öй²¾ÙÐÐÁË1273´ÎСRNA²âÐòʵÑé¡£ÔÚÒ»Ïîºá¶ÏÃæÑо¿ÖУ¬¶ÔÀ´×ÔÒâ´óÀûºÍ½Ý¿ËÐÐÁеķà±ãÑù±¾(155Àý½áÖ±³¦°©£¬87ÀýÏÙÁö£¬96ÀýÆäËû³¦µÀ¼²²¡£¬141Àý½á³¦¾µÒõÐÔ±ÈÕÕ)µÄmiRNAÆ×¾ÙÐÐÁËÑо¿¡£25¸ömiRNAÔÚCRC»¼Õߵķà±ãÖÐÏÔʾ¸Ä±äµÄˮƽ£¬°üÀ¨miR-149-3p¡¢miR-607-5p¡¢miR-1246¡¢miR4488ºÍmiR-6777-5pÔÚÄÚµÄÎåÖÖmiRNAÌØÕ÷½«»¼ÕßÓë±ÈÕÕ×éÇøÍÑÀëÀ´£¬²¢ÔÚ×ÔÁ¦ÐÐÁÐÖлñµÃÑéÖ¤¡£¸ÃÌØÕ÷½«±ÈÕÕ×é´ÓµÍ/¸ß·ÖÆÚÖ×ÁöºÍÍíÆÚÏÙÁöÖÐÇø·Ö³öÀ´(AUC=0.82, 95% CI=0.71-0.97)¡£×éÖ¯miRNAÆ×·´Ó¦ÁË·à±ãÑù±¾µÄmiRNAÆ×£¬¶ø²î±ð賦µÀ¼²²¡µÄ·à±ãÆ×Í»³öÁËCRCÖÐÌØÒìÐÔʧµ÷µÄmiRNA¡£FITÊ£ÓàÑùÆ·ÖеÄmiRNAÆ×ÓëÉúÑÄ»º³åÒºÖÐÍøÂçµÄ·à±ãµÄmiRNAÆ×¾ßÓÐÓÅÒìµÄÏà¹ØÐÔ£¬²¢ÇÒÔÚÏÙÁö»òCRC»¼ÕßÖпÉÒÔ¼ì²âµ½ËüÃǵĸı䡣